Bioprinting of Stem Cells Interplay of Bioprinting Process, Bioinks and Stem Cell Properties

Subscriber access provided by TUFTS UNIV
Review
Bioprinting of stem cells: interplay of bioprinting
process, bioinks and stem cell properties.
Supeng Ding, Lu Feng, Jiayang Wu, Fei Zhu, Ze'en Tan, and Rui Yao
ACS Biomater. Sci. Eng., Just Accepted Manuscript • DOI: 10.1021/acsbiomaterials.8b00399 • Publication Date (Web): 25 Jul 2018
Downloaded from http://pubs.acs.org on July 27, 2018
Just Accepted
“Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted
online prior to technical editing, formatting for publication and author proofing. The American Chemical
Society provides “Just Accepted” as a service to the research community to expedite the dissemination
of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts appear in
full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been fully
peer reviewed, but should not be considered the official version of record. They are citable by the
Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered to authors. Therefore,
the “Just Accepted” Web site may not include all articles that will be published in the journal. After
a manuscript is technically edited and formatted, it will be removed from the “Just Accepted” Web
site and published as an ASAP article. Note that technical editing may introduce minor changes
to the manuscript text and/or graphics which could affect content, and all legal disclaimers and
ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors or
consequences arising from the use of information contained in these “Just Accepted” manuscripts.
is published by the American Chemical Society. 1155 Sixteenth Street N.W.,

Washington, DC 20036
Published by American Chemical Society. Copyright © American Chemical Society.
However, no copyright claim is made to original U.S. Government works, or works
produced by employees of any Commonwealth realm Crown government in the course
of their duties.
Page 1 of 75 ACS Biomaterials Science & Engineering
1
2
3
4 Bioprinting of stem cells: interplay of bioprinting process, bioinks
5
6 and stem cell properties
7
8
Supeng Ding 1,2, Lu Feng1, Jiayang Wu 1,3, Fei Zhu 1, Ze’en Tan 1, Rui Yao 1*
9
10
11
12
13 1 Department of Mechanical Engineering, Biomanufacturing and Rapid Forming Technology
14
15
16 Key Laboratory of Beijing, Tsinghua University, Beijing, People’s Republic of China
17
18 2 Department of Materials Science and Engineering, Tsinghua University, Beijing, People’s
19
20
21 Republic of China
22
23 3 Department of Construction Management, Tsinghua University, Beijing, People’s Republic
24
25
26 of China
27
28
29
30
31
32
33 Corresponding Author*:
34
35 Rui Yao: yaorui@tsinghua.edu.cn
36
37
38
39
40
41
42
43 Abstract
44
45 Combining the advantages of 3D bioprinting technology and biological characteristics of stem
46
47
48 cells, bioprinting of stem cells is recognized as a novel technology with broad applications in
49
50 biological study, drug testing, tissue engineering and regenerative medicine, etc. However, the
51
52
biological performance and functional reconstruction of stem cells are greatly influenced by
53
54
55 both the bioprinting process and post-bioprinting culture condition, which are critical factors
56
57
1
58
59
60 ACS Paragon Plus Environment
ACS Biomaterials Science & Engineering Page 2 of 75
1
2
3
4 to consider for further applications. Here we review the recent development of stem cell
5
6 bioprinting technology and conclude major factors regulating stem cell viability, proliferation,
7
8
differentiation and function from the aspects of the choice of bioprinting techniques, the
9
10
11 modulation of bioprinting parameters and the regulation of stem cell niche in the whole
12
13 lifespan of bioprinting practice. We aim to provide a comprehensive consideration and
14
15
16 guidance of bioprinting of stem cells for optimization of this promising technology in
17
18 biological and medical application.
19
20
21
22
23 Keywords: 3D Printing, Biofabrication, Multipotent Stem Cells, Pluripotent Stem Cells, Stem
24
25
26 Cell Niche Regulation, Long-term Culture
27
28
29
30
31
32
33 1. Introduction
34
35 It has become a consensus that research on stem cells is of great significance in medicine,
36
37
38 physiology and biology study since they demonstrate unique abilities of proliferation and
39
40 differentiation compared with mature primary cells. Based on these features, stem cell
41
42
43 technology has been widely applied in the treatment of disease 1,2, drug development 2,3,
44
45 transgenic production 4, the pathological study 5, and has revealed potentials in organ
46
47
48 transplantation and regenerative medicine 2, etc. Nevertheless, being sensitive to the
49
50 microenvironment, stem cell properties can be influenced by many physical and chemical
51
52
factors, causing low viability, limited proliferation and undesirable differentiation in many
53
54
55 cases 1,4,6-8. This has become a primary obstacle in the development and application of stem
56
57
2
58
59
1
2
3
4 cell technology. Therefore, there is a pressing need for technologies offering specific design
5
6 of stem cell niche with the mild procedure to satisfy both the essential biological requirements
7
8
of stem cells and directed differentiation to desired lineages.
9
10
11
12
13 Bioprinting technology is commonly known as the process of creating cell and biomaterial
14
15
16 patterns in a confined space using 3D printing technique, where cell function and viability are
17
18 preserved within the printed construct 9,10. To realize transition from printable fluid to
19
20
21 long-term stable structure, cells are encapsulated within bioinks, which refer to biomaterials
22
23 with the ability to be deposited by certain bioprinting techniques and providing the
24
25
26 microenvironment for cell adhesion, proliferation and differentiation after bioprinting 11. In
27
28 2015, “bioprinting” became an official term in the Oxford Dictionary. It was defined as the
29
30
use of 3D printing technology with materials that incorporate viable living cells, e.g. to
31
32
33 produce tissue for reconstructive surgery. This technology can serve as one of the solutions
34
35 for the construction of geometrically, physically and chemically controlled stem cell
36
37
38 microenvironment. Compared with traditional methods for constructing 3D cell-laden
39
40 structures, such as hanging-drop 12, multi-well plate 13, and stamping 14, bioprinting
41
42
43 technology has an irreplaceable advantage of building delicate, complex and well-designed
44
45 microstructure.
46
47
48
49
50 Bioprinting process is applicable for different purposes because of adjustable bioprinting
51
52
principle and parameters, including printing speed, pressure, resolution, etc 15. Various bioinks
53
54
55 can be used for inducing stem cell fate by offering physical and chemical cues 16,17, while
56
57
3
58
59
1
2
3
4 additional factors can be supplemented in the bioinks or culture media for further induction or
5
6 maturation 18,19. As a result, stem cell bioprinting has become the rational choice by many
7
8
researchers for either enhancing the understanding of stem cells or enlarging the range of
9
10
11 application of both stem cell and bioprinting technology in tissue engineering, regenerative
12
13 medicine, drug discovery, etc. According to the Pubmed database, 350 articles are available
14
15
16 by searching with the keywords ‘(bioprinting OR printing) AND stem AND cell’ in recent 5
17
18 years, with a trend of continuous growth in recent years, indicating the booming of this field.
19
20
21
22
23 However, both bioprinting process and subsequent culture condition might lead to unwanted
24
25
26 effects on stem cells properties. Several factors such as mechanical stress 20, working
27
28 temperature 21, radiation 22 and bioink properties 23-26 are associated with cell injury and
29
30
undesired differentiation when administrated improperly. Therefore, it is necessary to
31
32
33 emphasize understanding of the underlying interactions among bioprinting process, bioinks
34
35 and stem cells in the whole lifespan of stem cell bioprinting practice. Bioprinting process
36
37
38 should satisfy the requirement of fabrication as well as stem cells at the same time. Bioinks
39
40 should demonstrate printabilty in order to create delicate and stable structure, and also
41
42
43 biocompatibility to induce stem cell differentiation and support their functions. Focusing on
44
45 these interactions, we provide guidance and perspective about how to choose or even develop
46
47
48 suitable bioprinting techniques, parameters and bioinks depending on the stem cell types and
49
50 specific application.
51
52
53
54
55 Bioprinting factors related to stem cell behaviors are elucidated in three stages, namely:
56
57
4
58
59
1
2
3
4 before bioprinting, during bioprinting and after bioprinting. Before bioprinting, suitable
5
6 bioprinting techniques should be chosen based on types of stem cells, properties of the desired
7
8
structure, and anticipated cost. During bioprinting, printing parameters ought to be optimized
9
10
11 to mitigate harm to stem cells. After bioprinting, microenvironment created by bioinks and
12
13 culture media can further regulate long-term cell proliferation and differentiation (Figure 1).
14
15
16 Since bioinks play the leading role in establishment of delicate structure and creation of
17
18 biocompatible microenvironment for stem cell culture, they are discussed specifically. We
19
20
21 overview recent achievements in the whole lifespan of stem cell bioprinting and discuss
22
23 potential challenges and improvements for future research.
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46 Figure 1. Schematic of the whole lifespan of stem cell bioprinting practice. Reproduced with
47
48
49 permission from ref 27, 28. Copyright 2015 and 2016 Biofabrication.
50
51
52
53
54 2. Choosing suitable bioprinting techniques
55
56 In order to satisfy demands of various research, different bioprinting techniques have been
57
5
58
59
1
2
3
4 developed from perspectives of manufacturing abilities (printing resolution, speed, throughput,
5
6 etc.) 29 and cell requirements (cell viability, proliferation, differentiation, etc.) 14.
7
8
There are two ways of classifying bioprinting technology, which are not mutually exclusive.
9
10
11 One way is based on the cell format (dispersed individual cells or cell aggregates) and cell
12
13 number resulted from each printing motion (only one cell or more than one cell), bioprinting
14
15
16 techniques can be mainly divided into multi-cell, single-cell and cell aggregate bioprinting. In
17
18 multi-cell bioprinting, large quantities of dispersed individual cells are dispersed in bioinks
19
20
21 and more than one cell is delivered within a droplet or a filament. In single-cell bioprinting,
22
23 merely one cell is printed at each printing motion. In cell aggregate bioprinting, pre-formed
24
25
26 cell aggregates instead of dispersed individual cells are embedded in the bioinks and then
27
28 bioprinted.
29
30
31
32
33 Another method to classify bioprinting techniques is based on enabling technology, which are
34
35 commonly seen in current reports, namely inkjet bioprinting, microextrusion-based
36
37
38 bioprinting, laser-assisted bioprinting and stereolithography-based bioprinting. We choose to
39
40 introduce the four enabling techniques under multi-cell bioprinting category, since compared
41
42
43 with single-cell bioprinting and cell aggregate bioprinting, it is the most mature and
44
45 developed field (Figure 2).
46
47
48
49
50
51
52
53
54
55
56
57
6
58
59
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46 Figure 2. Illustrations and examples of different bioprinting techniques. (A) Multi-cell bioprinting,
47
48
49 which can be further divided into inkjet bioprinting (image from MicroFab Technologies, Inc.),
50
51 microextrusion-based bioprinting (image from Organovo Holdings, Inc.), laser-assisted bioprinting
52
53
54 (image from Organovo Holdings, Inc.) and stereolithography-based bioprinting (image from Formlabs,
55
56 Inc.). (B) Cell aggregate bioprinting. Reproduced with permission from ref 30. Copyright 2009
57
7
58
59
1
2
3
Biomaterials. (C) Single-cell bioprinting. Reproduced with permission from ref 31. Copyright 2012
4
5
6 Biofabrication.
7
8
9
10
11 2.1. Multi-cell bioprinting
12
13 2.1.1. Inkjet bioprinting
14
15
16 Inkjet bioprinting of cells was first reported by Roth E. A. et al. in 2004 to achieve
17
18 high-throughput patterning of smooth muscle cells 32. Also known as drop-on-demand
19
20
21 bioprinting, it is defined as dispensing through a small orifice and precise positioning of small
22
23 volumes (1–100 picolitres) of bioink and/or cells on a substrate to fabricate 3D cell patterns or
24
25
26 3D cell-laden constructs 33. Thermal 34, piezoelectric 35, valve-based 36 and electric field-based
27
28 inkjet bioprinting 37,38 are the most commonly adopted approaches. Thermal inkjet bioprinters
29
30
function by heating the printing head to produce pulses of pressure that forces droplets out of
31
32
33 the nozzle. In piezoelectric inkjet bioprinting, a mechanical pulse is applied to the fluid in the
34
35 nozzle by a piezoelectric actuator causing a shock wave that forces the bioink throughout the
36
37
38 nozzle. In electric field-based inkjet bioprinting, bioinks are forced to leave the nozzle by
39
40 static electric force and then form a droplet in the crosslinking medium. Inkjet bioprinting
41
42
43 demonstrates a series of advantages including low cost, high resolution, high printing speed
44
45 and high post-bioprinting viability and has been employed on stem cells in recent years.
46
47
48 Multipotent stem cells including mesenchymal stem cells (MSC) 39,40, muscle-derived stem
49
50 cells 41, and neural stem cells (NSC) 42 have been bioprinted for regenerative medicine
51
52
purposes. In addition, co-printing of amniotic fluid-derived stem cells and canine smooth
53
54
55 muscle cells was reported by Xu T. et al., demonstrating the feasibility of establishing
56
57
8
58
59
1
2
3
4 heterogeneous tissue structure from stem cells by inkjet bioprinting technology 43. As for
5
6 pluripotent stem cells, Faulkner-Jones A. et al. bioprinted embryonic stem cells (ESC) and
7
8
induced pluripotent stem cells (iPSC) to study stem cell differentiation in 3D
9
10
11 microenvironment 21.
12
13
14
15
16 Physical and chemical factors involved in bioprinting process, including mechanical force,
17
18 bioink viscosity and temperature should be paid attention to in the practice of inkjet
19
20
21 bioprinting of stem cells. Stem cell injury and even death may result from both the shear
22
23 stress by extrusion through small orifices of the print head 20 and/or the normal stress by
24
25
26 reaching the substrate. Besides, bioinks used in inkjet bioprinting are confined to low
27
28 viscosity to avoid frequent clogging. However, low viscosity bioink usually ends up with
29
30
immediate droplet spreading on the substrate, impeding the building of designed 3D
31
32
33 constructs with sound shape fidelity and mechanical stability 22. What’s more, the high
34
35 temperature in thermal inkjet bioprinting can be a potential risk, although the duration of
36
37
38 localized heating is short (~2µs) and little harm to cells has been reported 44. Xu T. et al.
39
40 reported that thermal inkjet bioprinting could have a benign effect on simultaneous gene
41
42
43 transfection by co-printing of porcine aortic endothelial cells (pAEC) and plasmids encoding
44
45 green fluorescent protein 45. Further research on the influence of high temperature on cell
46
47
48 biology and function in thermal inkjet bioprinting is still required.
49
50
51
52
2.1.2. Microextrusion-based bioprinting
53
54
55 Microextrusion-based bioprinting, also known as bioplotting technology, was first reported by
56
57
9
58
59
1
2
3
4 Fedorovich N. E. et al. in 2007 that spatially organized constructs could be fabricated by
5
6 microextrusion-based bioprinting of bone marrow stromal cell-laden alginate 46. In this
7
8
technique, continuous filaments rather than liquid droplets of bioinks are extruded throughout
9
10
11 a micro-nozzle to form 3D structures 15. One of the outstanding features is that the viscosity
12
13 of bioinks used can range from 30 mPa·s to more than 6 × 107 mPa·s, providing much
14
15
16 broader bioink options to meet requirements of different types of stem cells and provide
17
18 desired mechanical strength 47. Besides, shear-thinning bioinks, which exhibit the
19
20
21 non-Newtonian behavior of fluids whose viscosity decreases under shear strain, have been
22
23 explored to further improve the printing resolution and shape fidelity of the 3D constructs 48.
24
25
26 Other advantages includes high cell density 22 and high printing throughput, which enables
27
28 fabrication of organized constructs with clinically relevant sizes 49,50. bioprinting is also
29
30
recommended as an inexpensive bioprinting approach for industrial application in future,
31
32
33 where Reid J. A. et al. invented a 3D printer with a cost less than $200 for iPSC bioprinting
34
35 51
.
36
37
38
39
40 In recent years, further optimization has been developed in this technique. Co-axial extrusion
41
42
43 of multi-components with a microfluidic bioprinting head has been realized for reaction-based
44
45 bioink crosslinking. Since only the surface of the filament is crosslinked and the inner bioinks
46
47
48 remain liquid during bioprinting process, a core-shell structure of filament is generated, where
49
50 the surface provides a good formability and the inner part with low viscosity ensures high cell
51
52
viability 52,53,54. Multi-nozzle bioprinters can improve printing throughput and support
53
54
55 deposition of multi-bioinks encapsulated with various cells, holding the potential of
56
57
10
58
59
1
2
3
4 regenerating complex tissues 55,56. Moreover, Highley C. B. et al. explored a method to create
5
6 constructs with complex structure by bioprinting a second bioink inside a support self-healing
7
8
hydrogel 57. Based on its shear-thinning property, the self-healing hydrogel could deform
9
10
11 when a syringe needle was inserted, but recovered rapidly to retain the printed structure. As
12
13 the most common and affordable bioprinting techniques, bioprinting has been widely used in
14
15
16 printing multipotent stem cells including bone marrow-derived mesenchymal stem cells
17
18 (BMSC) 58, adipose-derived stem cells (ASC) 59,60, neural stem cells 61,62, and glioma stem
19
20
21 cells 63. In recent years, more and more research has been focused on pluripotent stem cells.
22
23 For example, Ouyang L. et al. utilized bioprinting technique to fabricate ESC-laden hydrogel
24
25
26 structure with good printability and high cell viability, where printing temperature, holding
27
28 time and bioink concentration can be controlled precisely 27. Moreover, it was reported by Gu
29
30
Q. et al. that bioprinting of iPSCs could facilitate cell proliferation and successive
31
32
33 differentiation 64.
34
35
36
37
38 Although widely used, several limitations need to be aware of in bioprinting research. Firstly,
39
40 high bioink viscosity may reduce cell survival rate, mainly attributed to the shear stress
41
42
43 inflicted on cells in viscous fluids 27,65. Secondly, extrusion pressure should be gentle enough
44
45 to avoid increased cell death, which restricts further improvement of bioprinting speed and
46
47
48 throughput. Thirdly, a decrease of nozzle size to achieve higher printing resolution can affect
49
50 cell survival as well 66. Therefore, bioprinting process and parameters need to be carefully
51
52
adjusted to achieve both high cell viability and sound printability.
53
54
55
56
57
11
58
59
1
2
3
4 2.1.3. Laser-assisted bioprinting
5
6 Laser-assisted bioprinting was first proposed by Ringeisen B. R. et al. in 2004, where
7
8
pluripotent murine embryonal carcinoma cells embedded in matrigel were transferred to the
9
10
11 substrate in an organized pattern with little cell damage 67. Based on the principles of
12
13 laser-induced forward transfer (LIFT), laser pulses are focused on an energy-absorbing layer
14
15
16 to provide energy for generating bubbles in another layer of cell-laden bioinks, which
17
18 subsequently propel bioinks from the donor film to the receiving substrate 15,68,69. As a
19
20
21 nozzle-free method, problems including bioink clogging and cell death resulted from shear
22
23 stress and can be avoided, leading to higher cell viability. Besides, higher printing resolution
24
25
26 can be achieved in this techniques, which is dependent on bioink viscosity, laser energy and
27
28 scanning speed 70. Ali M. et al. reported laser-assisted bioprinting of MSCs with post-printing
29
30
cell viability close to 100% 71. Sorkio A. et al. reported stroma-mimicking structures could be
31
32
33 generated by laser-assisted bioprinting of ADSCs 72. Dias A. D. et al. investigated
34
35 laser-assisted bioprinting of ESC, which served as an approach for the formation of the
36
37
38 embryoid body 73.
39
40
41
42
43 Nevertheless, laser-assisted bioprinting is relatively time-consuming, rendering limitations in
44
45 the high-throughput study since overlong printing time can reduce cell viability 16. Moreover,
46
47
48 the cost of establishing laser-assisted bioprinting system is much higher than the above two
49
50 techniques. Therefore, this technique is suggested to be applied in the fabrication of delicate
51
52
structure of small size.
53
54
55
56
57
12
58
59
1
2
3
4 2.1.4. Stereolithography-based bioprinting
5
6 Stereolithography-based bioprinting was first used for establishment of human dermal
7
8
fibroblasts-laden poly(ethylene glycol) (PEG) structure by Arcaute K. et al. in 2006 74. Liquid
9
10
11 biocompatible resins are solidified by photopolymerization under irradiation in a designed
12
13 pattern to fabricate layer-by-layer construct 75. As another nozzle-free technique, mechanic
14
15
16 shear stress can be avoided. High bioprinting resolution is a noteworthy advantage where
17
18 commercial systems usually create patterns with the resolution around 100µm~200µm. The
19
20
21 resolution can even reach 50µm, in a case of bioprinting 3T3 fibroblast cells encapsulated in
22
23 the mixture of polyethylene glycol diacrylate (PEGDA) and gelatin methacrylate (GelMA)
24
25
26 hydrogel 76. Since crosslinking is realized by covalent binding, another advantage of
27
28 stereolithography-based bioprinting is the long-term stability. As a consequence, constructs
29
30
with fine pore structure can be fabricated, which is significant in the maintenance of
31
32
33 mechanical properties and mass transfer for long-term cell culture 75. In stem cell research,
34
35 stereolithography-based bioprinting has been employed in bioprinting multipotent stem cells
36
37
38 such as BMSCs 77-80, ASCs 81, and NSCs 82, for enhanced differentiation or tissue repair. For
39
40 pluripotent stem cells, Bajaj P. et al. reported that ESC encapsulated in PEGDA could be
41
42
43 patterned by stereolithography-based bioprinting for 3D culture 83.
44
45
46
47
48 Bioinks utilized in Stereolithography-based bioprinting technology need to be biocompatible,
49
50 biodegradable and photocurable, resulting in relatively limited choice. Commonly used
51
52
bioinks includes PEGDA, poly(propylene fumarate) (PPF), PTMC, poly(e-caprolactone)
53
54
55 (PCL), GelMA, and poly(D,L-lactide) (PDLLA). Combination of photocurable materials and
56
57
13
58
59
1
2
3
4 natural ECM matrix is also a common option to improve cell performance and/or induce
5
6 desired differentiation and function 75,76,84. But it is reported that the long-term cell viability in
7
8
stereolithography-based bioprinting is lower than the techniques above, since these bioinks
9
10
11 have less biocompatibility and photoinitiators may decrease cell survival 85. Besides, the
12
13 energy of UV light may result in cell death or induce carcinogenesis to some extent, which
14
15
16 can be more serious in bioprinting of stem cells 76. Thus, the development of visible light
17
18 crosslinking bioinks is a clear trend to broaden the perspective of this promising technology.
19
20
21
22
23 2.2. Cell aggregate bioprinting
24
25
26 Unlike multi-cell bioprinting where single cells are individually distributed in the bioink, cell
27
28 aggregate bioprinting utilizes the properties of cell aggregates to mimic the structure and
29
30
function of tissues in vivo such as organoids 86. Cells in aggregates experience a divergent
31
32
33 microenvironment comparing with single cells, since cell-cell contacts increase and
34
35 cell-substrate contacts decrease. Coburn L. et al. modeled the cellular force distribution inside
36
37
38 an epithelial cell aggregate and demonstrated that contact inhibition of locomotion can
39
40 modulate the mechanical response of cells 87. In addition, immunomodulatory paracrine factor
41
42
43 secretion increased in MSC aggregates due to the enhancement of intercellular adhesion and
44
45 cell contact-dependent signaling 88. Komatsu M. et al. demonstrated that aggregates can assist
46
47
48 maturation of dopamine neuron precursors into neurons 89. Meanwhile, cell aggregate
49
50 bioprinting has a much higher throughput, since the number of cells in an aggregate
51
52
commonly ranges from 500 to 250,000 90. Therefore, this technique is of significance for
53
54
55 tissue regeneration of larger scale.
56
57
14
58
59
1
2
3
4
5
6 The process for cell aggregate bioprinting includes the formation of cell aggregates,
7
8
bioprinting of aggregates, and aggregate fusion and maturation. With self-assembly and
9
10
11 self-organization, generation of cell aggregates can be realized by several methods, such as
12
13 hanging-drop culture, non-adhesive surface, scaffolds, and microfabrication 91, with different
14
15
16 aggregate shapes like spheroids, cylinders 92 and sheets 49. These cell aggregates should have
17
18 the adequate mechanical strength to avoid fracture during bioprinting and long-term culture,
19
20
21 which is an essential problem when considering the type of cells for cell aggregate
22
23 bioprinting.
24
25
26
27
28 Subsequently, cell aggregates are printed by specific approaches, which can be mainly
29
30
divided into two types based on cell-substrate contact. The first one is similar to multi-cell
31
32
33 bioprinting where cell aggregates are encapsulated in bioactive bioinks, which provide
34
35 adhesion ligands. For example, Marchioli G. et al. bioprinted islets encapsulated in a mixture
36
37
38 of gelatin and alginate for islet transplantation 93. In the second one, namely scaffold-free
39
40 bioprinting, bioinert materials like alginate 94 or agarose 31 and cell aggregates are co-printed.
41
42
43 Cell aggregates are deposited adjacently without the barrier of materials and bioinks merely
44
45 function as upholders before the cell aggregates complete the fusion, forming a tissue stable
46
47
48 enough to hold itself.
49
50
51
52
After bioprinting, a quick fusion of cell aggregates is important for building stable tissues.
53
54
55 Since cell aggregates serve as building blocks in the structure, the stability of the printed
56
57
15
58
59
1
2
3
4 structure can be enhanced via aggregate fusion by improving inter-aggregate force 92. This
5
6 process is suggested to be regulated by cellular interaction and cell migration, and in some
7
8
case analyzed by phase field theory 95,96. Maturation of cell aggregates is another vital process.
9
10
11 To mimic the tissue formation in vivo, additional factors can be considered to modulate
12
13 microenvironment of cell aggregates, accelerating maturation to generate stable and
14
15
16 functional tissues 97.
17
18
19
20
21 2.3. Single-cell bioprinting
22
23 Single-cell bioprinting aims to distribute singular cell in a controlled way for the fabrication
24
25
26 of delicate tissues and single cell studies, such as genetic characterization of single cells 98,99.
27
28 It is especially significant for stem cell research since the cells and microenvironment
29
30
components can be accurately arranged.
31
32
33
34
35 Such an idea was first proposed in 2005 just after the invention of laser-assisted bioprinting,
36
37
38 as Barron J. A. et al. reported laser-assisted bioprinting of human osteosarcoma cells. They
39
40 utilized bioinks with different cell concentration to control the cell numbers in one droplet 100.
41
42
43 However, although the size of droplets was homogeneous and small enough, one cell in each
44
45 droplet was hardly realized and the cell number in one droplet basically obeyed Poisson
46
47
48 distribution whatever the cell concentration was 100. Therefore, it was until 2012 that the real
49
50 single-cell bioprinting had been invented, where the high-speed camera was used to check the
51
52
cell status in the ejection area of the nozzle tip. When there was no cell or over one cell, the
53
54
55 droplet was discarded 32. In 2014, a block-cell-printing technique was proposed by Zhang K.
56
57
16
58
59
1
2
3
4 et al., where hook-shaped traps were positioned in designated patterns to trap singular breast
5
6 cancer cell and work as printing nozzle 101. Although single-cell bioprinting of stem cells has
7
8
not been reported as far as we know, there is a clear trend in the emerging of stem cell
9
10
11 bioprinting technology assisted single cell study, due to the significance of stem cell biology.
12
13
14
15
16 In conclusion, to choose a suitable bioprinting technique, both fabrication process and cell
17
18 requirements should be considered. Cell aggregate bioprinting is usually required in
19
20
21 generation of tissues with a tight junction between cells and of great size. With the ability to
22
23 build delicate structures, single-cell bioprinting is utilized in research on cellular-matrix
24
25
26 interaction by depositing stem cells and microenvironment elements with precisely defined
27
28 spatial location. On the other hand, multi-cell bioprinting is most widely used and most
29
30
developed technology. Inkjet bioprinting and microextrusion-based bioprinting are cost
31
32
33 efficient approach suitable for relatively big and rough structures while preserving high cell
34
35 viability, while laser-assisted bioprinting can offer high resolution with little cell death.
36
37
38 Stereolithography-based bioprinting can generate even more elaborate structures, but is less
39
40 biocompatible. In addition, it is better to choose a technique with lower cost and gelation
41
42
43 methods suitable for the specific lab equipment. In all, after a comprehensive consideration of
44
45 the size, precision and complexity of desired structures and the type of cells, limitations can
46
47
48 be determined from various aspects including printing speed, throughput, resolution, cell
49
50 viability and density, based on which we can decide the suitable bioprinting technique. Table
51
52
1 can be used to help with choice of bioprinting technique.
53
54
55
56
57
17
58
59
1
2
3
4
5
6
7
8
Table 1. Comparison of bioprinting techniques
9
10
11 Multi-cell bioprinting
Cell aggregate
12 Techniques
Inkjet bioprinting
Microextrusion-based Laser-assisted Stereolithography-based bioprinting
Single-cell bioprinting
13 bioprinting bioprinting bioprinting
14 Printing speed High Low Medium High Medium High
15 Printing
Medium High Medium High High Low
throughput
16
Resolution Medium Low High High Low High
17
18 Cell viability Medium Medium High Low High Medium
19 Cell density Low High Medium Medium High Low
20 Cost Low Low High High Medium High

21
Ionically, Shear-thinning;
22 thermal, ionically, Ionically,
Shear-thinning;
23 Crosslinking
methods
pH mediated;
polymerized,
thermal,
pH mediated;
thermal;
photopolymerized;
Photopolymerized ionically,
thermal
Ionically,
thermal
24 photopolymerized;
enzymatic
polymerized,
photopolymerized
enzymatic
25 References [20,22,34,35,44,102] [27,47-50,65,66,103] [16,70,71,104] [75,76,94,95] [86,90,94] [32,101,105,106]

26
27
28
29
30
31
3. Bioprinting process regulating stem cell properties
32
33 In bioprinting process, stem cells will experience a period of transfer from a reservoir to an
34
35 arranged position, during which external factors, e.g. mechanical stress, heat, radiation may
36
37
38 be involved. These factors will inevitably influence stem cell properties Therefore, a
39
40 systematic review of the interplay between stem cells and bioprinting process is necessary to
41
42
43 customize bioprinting techniques for stem cells with regulated performance in subsequent
44
45 culture.
46
47
48
49
50 3.1. Mechanical stress
51
52
Mechanical stress is requisite to drive the cells and bioinks to the platform in any bioprinting
53
54
55 techniques, leading to deteriorating of cell properties. Although the biochemical process of
56
57
18
58
59
1
2
3
4 cell death caused by external force is still unclear, excessive deformation of cells under
5
6 internal strain is reported as a phenomenological reason 107. As for stem cells, differentiation
7
8
can be impacted by stress as well, since mechanical force can act on mechanoreceptor
9
10
11 integrins, pericellular tethers, focal adhesions ion channels, cadherins, connexins, and the
12
13 plasma membrane’s lipid rafts 108.
14
15
16
17
18 Mechanical stress can be divided into normal stress and shear stress. Normal stress refers to
19
20
21 the stress perpendicular to the cell surface when bioinks are extruded through bioprinting
22
23 machine or landed on the substrate, while shear stress refers to the stress parallel to the cell
24
25
26 surface when viscous fluid bioinks flow through the nozzle. In nozzle-free bioprinting
27
28 techniques such as laser-assisted bioprinting or stereolithography-based bioprinting, merely
29
30
normal stress affects cells in most cases, thus an open-pool ejection system is suggested from
31
32
33 the point of reducing potential loss of cell viability 22. Shear stress is regarded as the major
34
35 cause of cell injury and death in bioprinting techniques with nozzles, especially
36
37
38 microextrusion-based bioprinting with viscous bioinks (Figure 3A). Higher shear stress is
39
40 generated by either greater driving force to improve printing speed and efficiency 21,109-111 or
41
42
43 smaller nozzle diameter to increase printing accuracy 112-114. Viscous bioinks may cause more
44
45 cell death due to increased shear stress, yet they can facilitate the shape fidelity of 3D
46
47
48 structure 22. Shear thinning bioinks are proposed as a solution to improve cell viability, since
49
50 their viscosity decreases as shear rate increases 22,115. As reported by Thakur A. et al.,
51
52
hydrogels can be modified with nanosilicates to enhance the shear-thinning characteristic for
53
54
55 the better viability of MSC after injection through a nozzle 116. Non-viscous bioinks that
56
57
19
58
59
1
2
3
4 solidify rapidly after leaving needles serve as another choice, which can be realized by
5
6 UV-light photo-crosslinking 117 or ionically crosslinking via coaxial extrusion 118. Jason
7
8
Burdick’s group invented an in situ photo-crosslinking approach for bioprinting low-viscous
9
10
11 cell-laden methacrylated hyaluronic acid through a photopermeable capillary to achieve
12
13 immediate crosslinking prior to deposition (Figure 3B) 117.
14
15
16
17
18 3.2. Temperature and radiation
19
20
21 Besides mechanical stress, temperature and radiation are other physical cues affecting stem
22
23 cell properties during bioprinting process. Compared with low temperature, cells are more
24
25
26 sensitive to high temperature, which can be resulted from localized heating in thermal inkjet
27
28 bioprinting or thermal energy of UV light in laser-assisted bioprinting (Figure 3C). Thus, the
29
30
regulation of temperature in the whole bioprinting process is significant in the design of
31
32
33 bioprinting techniques 120. It was reported that localized temperature up to 300oC did not
34
35 exhibit a substantial impact on the viability of the neural cells 121. As for laser-assisted
36
37
38 bioprinting, Yaakov N. et al. demonstrated using near-infrared wavelength (700–1000 nm)
39
40 could overcome the exposure of excessive thermal energy onto the cell biolayer and
41
42
43 overheating of the cell, since photons in the near-infrared lack the energy to generate free
44
45 radicals and are not absorbed by DNA 120. But moreover, radiation can be more dangerous in
46
47
48 laser-assisted bioprinting, stereolithography-based bioprinting or any photo-crosslinking
49
50 bioprinting techniques (Figure 3D). Schiele N. R. et al. demonstrated that excessive
51
52
absorption of radiation energy may lead to cell injury and DNA damage of pluripotent
53
54
55 embryonal mouse carcinoma cells 122, so the irradiation parameter ought to be monitored
56
57
20
58
59
1
2
3
4 carefully.
5
6 3.3. Duration of unfavorable conditions
7
8
During the whole process of bioprinting, cells experience an unsatisfying environment,
9
10
11 including insufficient culture media, harmful chemicals (e.g. crosslinking agent) and complex
12
13 physical cues. Therefore, the duration of unfavorable conditions is supposed to be shortened
14
15
16 as much as possible to increase cell viability. For example, a shorter nozzle is proposed in
17
18 bioprinting to reduce the extrusion time, when cells experience shear force 15. Cui X. et al.
19
20
21 recommend that the maximum heating duration of each cell in thermal inkjet bioprinting
22
23 should be limited to 2 µs 44. On the other hand, the duration has to be long enough to satisfy
24
25
26 some process requirements, such as sufficient crosslinking for the construction of stable
27
28 structures. In addition, loss of cell viability can also result from increased stress when printed
29
30
with high speed while pursuing short printing time. Therefore, the optimization of bioprinting
31
32
33 duration should be considered comprehensively.
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
21
58
59
1
2
3
Figure 3. Regulation of stem cell properties by controlling mechanical stress, temperature, and
4
5
6 radiation in bioprinting process. (A) Stress distribution in the needle during microextrusion-based
7
8
bioprinting. Comparing with static status, stress in flowing bioinks can lead to cell death near the shell
9
10
11 of the needle. Reproduced with permission from ref 109. Copyright 2016 Advanced Healthcare
12
13 Materials. (B) A structure generated by bioprinting of low-viscous methacrylated hyaluronic acid. In
14
15
16 situ crosslinking makes it feasible to bioprint low-viscous bioinks with high fidelity, which can
17
18 decrease shear stress in microextrusion-based bioprinting. Reproduced with permission from ref 117.
19
20
21 Copyright 2016 Advanced Materials. (C) Viability of Chinese hamster ovary cells after thermal inkjet
22
23 bioprinting. Heat in thermal inkjet bioprinting caused cell death, which is related to cell concentration.
24
25
Reproduced with permission from ref 44. Copyright 2010 Biotechnology & Bioengineering. (D)
26
27
28 Spreading and viability of MSCs influenced by radiation in laser-assisted bioprinting. Reproduced with
29
30
permission from ref 77. Copyright 2017 Scientific Reports.
31
32
33
34
35
36
4. Microenvironmental factors modulating stem cell functions in
37
38 subsequent long-term culture
39
40 After bioprinting, stem cell-laden structures will be cultured for further investigation. The
41
42
43 microenvironment constructed by cell printing technology involves many aspects, including
44
45 micro- and macro-structure, biophysical and biochemical properties of bioink and culture
46
47
48 medium, and mass transfer, etc. These microenvironment factors would vary over time and
49
50 interact with stem cells to modulate their viability, proliferation and differentiation.
51
52
53
54
55 4.1. Biophysical cues
56
57
22
58
59
1
2
3
4 4.1.1. Stability and biodegradability
5
6 In many cases, bioprinted constructs need to be cultured in long-term for downstream
7
8
applications. One of the most important issues is the survival and preservation of functionality
9
10
11 of the encapsulated cells within the hydrogel bioink over time 110, thus posing an essential
12
13 demand for structure stability. Besides the collapse of the entire construct, Müller M. et al.
14
15
16 found that micro breaks can also reduce cell viability because of the distortion of stress field.
17
18 Structure stability can be attributed mainly to the intrinsic properties of bioink and specific
19
20
21 culture condition. Native biomaterials are employed in bioprinting for their prominent
22
23 biocompatibility, while most of them lacks long-term mechanical strength 58. Hence,
24
25
26 multi-materials or inorganic additions are also investigated 123. As for synthetic biomaterials,
27
28 crosslinking is considered as a reliable solution to improve stability, with various choices of
29
30
crosslinking mechanisms and reagents 22,115. Service performance is also significant in
31
32
33 practical application, especially in vivo research or transplantation. In vivo condition will
34
35 accelerate the degradation of materials by enzyme activity or other chemical reactions,
36
37
38 leading to the loss of filler volume 124. Wang X. F. et al. reported that a low-viscous bioink
39
40 consisting of gelatin and alginate embedding ASCs exhibited irregular morphology and loss
41
42
43 of structure in vivo after osteogenic differentiation in vitro, which resulted in insufficient
44
45 volume to fill the gap and lose connection between printed cells and tissues around, reducing
46
47
48 the efficiency of bone regeneration 125.
49
50
51
52
But on the other hand, the bioinks should be replaced by extracellular matrix (ECM)
53
54
55 ultimately. The ideal bioink should demonstrate the change of mechanical property in the
56
57
23
58
59
1
2
3
4 whole lifespan of bioprinting practice. That is, to support delicate structure after bioprinting,
5
6 to maintain shape fidelity during cell culture, and to degrade gradually with the rate matching
7
8
tissue fusion and maturation. The degradation process is suggested to keep pace with the
9
10
11 production of ECM 15 and the proliferation of stem cells 126. For example, Rutz A. L. et al.
12
13 invented a series of tunable hydrogels by mixing gelatin, fibrinogen, PEG, reactive
14
15
16 group-modified PEG, and peptide amphiphile under different ratios, to alter construct
17
18 degradation in accordance with ECM production, improving the viability and function of
19
20
21 human dermal fibroblasts 127.
22
23
24
25
26 4.1.2. Stiffness
27
28 Besides mechanical stability, stiffness is also strongly related to cell behaviors in stem cell
29
30
bioprinting. Attachment to the matrix is realized via integrin-containing focal adhesions,
31
32
33 which links to stress fibers such as actin-myosin cytoskeleton inside cells. These fibers can
34
35 pull the matrix, sense resistance and provide mechanical signals to regulate cell spreading,
36
37
38 morphology and differentiation 128. It is reported that with the same culture medium, MSC
39
40 differentiation can be induced merely by matrix stiffness to the lineage of neural (0.1–1 kPa),
41
42
43 myogenic (8–17 kPa) or osteogenic (>34 kPa), where signaling mediators of
44
45 mechanotransduction like TAZ and YAP sense matrix stiffness and regulate MSC
46
47
48 differentiation 24,129. Duarte Campos D. F. et al. has utilized this finding in
49
50 microextrusion-based bioprinting of MSC, and demonstrated enhanced differentiation
51
52
towards osteogenic in soft collagen-rich bioink and towards adipogenic in stiff agarose-rich
53
54
55 bioink 129 (Figure 4A).
56
57
24
58
59
1
2
3
4
5
6 4.1.3. Surface features
7
8
Since the reaction between stem cells and bioinks merely exists on the material surface where
9
10
11 cells attach, stem cell functions can be regulated by changing the surface features. Besides
12
13 chemical synthesis or coating to introduce ligands which are benefit to cell attachment,
14
15
16 survival or inducing desired differentiation, surface topography in nano- and micro-scale has
17
18 a significant effect on cell behavior such as cell growth, morphology 130, migration,
19
20
21 proliferation 131, differentiation 25 and cytoskeletal assembly 15 as well (Figure 4B). However,
22
23 most research is still at the stage of 2D surface patterning with micro-topography 132. Surface
24
25
26 modification of metal can be realized by acid etching, hydrothermal treatment and micro-arc
27
28 oxidation 130, while topographical surface on polymers can be generated by photolithography
29
30
and micromolding 132. Pioneer techniques have been developed in surface topography of 3D
31
32
33 structure as well, where Poellmann M. J. et al. fabricated microscale polyacrylamide (PAM)
34
35 on a second hydrogel by electrohydrodynamic jet bioprinting, where the behavior of
36
37
38 MC3T3-E1 preosteoblast cells was regulated by the size of PAM droplets 133. In the future,
39
40 bioprinting with higher resolution holds the potential to produce constructs with delicate
41
42
43 topographic features from micro-pores to nano-structures to optimize extracellular
44
45 environment.
46
47
48
49
50 Nowadays, mechanical and topological features have been widely applied in regulating stem
51
52
cell proliferation and differentiation. Actually, other physical signals can be utilized to
53
54
55 modulate post-bioprinting culture as well. For example, electrical stimulation was reported to
56
57
25
58
59
1
2
3
4 enhance neurogenesis of neural stem progenitor cells, neurite extension and neuron
5
6 maturation 134. In the future, the creation of a comprehensive physical field around printed
7
8
structures can be considered to enhance stem cell function and maturation.
9
10
11 4.2. Biochemical cues
12
13 4.2.1. Bioactive bioinks
14
15
16 Besides biocompatibility to ensure cell viability, traditional bioinks are mostly inertia to cell
17
18 differentiation to avoid undesired fate. However, since bioink functions as the major
19
20
21 component of stem cell niche, bioactive bioinks have been explored recently to facilitate stem
22
23 cell proliferation 135 or induce differentiation 26. Most bioactive bioinks are native materials or
24
25
26 the mixture of native and synthetic materials, as biospecific signals are presented in their
27
28 protein structure 136,137 (Figure 4C). For example, decellularized extracellular matrix (dECM)
29
30
has been widely used in stem cell bioprinting where cells encapsulated exhibit intrinsic
31
32
33 morphology, high viability, better proliferation and differentiation in vivo 26,138,139. Pati F. et al.
34
35 reported that the engraftment, survival and long-term function of adipose-derived stem cells
36
37
38 (ASC) and turbinate-tissue derived stromal cells were improved due to the crucial cues from
39
40 tissue-specific dECM bioinks. As for differentiation, cells were induced for adipogenic
41
42
43 differentiation by adipose dECM, chondrogenic differentiation by cartilage dECM and
44
45 cardiogenic differentiation by heart dECM 137. Polypeptide-DNA hydrogel, as another
46
47
48 example of bioactive bioinks, were invented by Li C. et al. for bioprinting of AtT-20 cells.
49
50 Composed of polypeptide and DNA, this material is biodegradable and programmable under
51
52
physiological conditions of proteases and nucleases 140.
53
54
55
56
57
26
58
59
1
2
3
4 4.2.2. Intercellular signals
5
6 Besides the cues from bioinks, signals released by other cells also direct stem cell behavior.
7
8
Intercellular signals can be divided into intracrine, autocrine, juxtacrine, paracrine, and
9
10
11 endocrine signals based on the distance between signaling and responder cells. In stem cell
12
13 bioprinting, juxtacrine signaling and paracrine signaling are more significant due to the size
14
15
16 of printed constructs nowadays. As for the former, it has been proved that intercellular
17
18 communication can be realized by molecule transfer through gap junction channels along the
19
20
21 contact surface 141. Stem cells which tend to form aggregates are influenced deeply by cluster
22
23 situation under this mechanism. The differentiated degree of MSC into osteogenic or
24
25
26 adipogenic lineage is reported much higher in aggregated cells than in isolated cells 142.
27
28 Besides, the size of stem cell aggregates can regulate differentiation fate 14. With the control
29
30
of the initial cell seeding density, bioprinting volume and culture time, stem cell aggregates
31
32
33 can be formed with a regular shape and uniform size through cell proliferation after
34
35 bioprinting 143. Ouyang L. et al. developed a method for mass production of embryoid bodies
36
37
38 through bioprinting technology, where ESC pluripotency was maintained with optimized
39
40 aggregate size and morphology 144. Contact intercellular signals also exist in cell aggregate
41
42
43 bioprinting where aggregates are fabricated in advance, as Wang Z. et al. reported enhanced
44
45 functionalization of MSC aggregates in printed gelatin-based structure, comparing with single
46
47
48 MSCs under the same bioprinting condition 145.
49
50
51
52
Paracrine signals are released by signaling cells and target the cells in the vicinity. One of the
53
54
55 most important employment is co-culture of multiple cells to improve cell functions. Roy N.
56
57
27
58
59
1
2
3
4 S. et al. reported that differentiation of human ESCs towards dopaminergic neuronal fate can
5
6 be enhanced by co-culture with human fetal midbrain astrocytes 146. Yao R. et al. found
7
8
co-culture of ASC and human umbilical vein endothelial cells (HUVEC) encapsulated in
9
10
11 collagen/alginate microsphere could facilitate in vivo vascularization and formation of
12
13 adipose tissues 147. Co-printing of different types of cells to control cell migration has been
14
15
16 studied as well, where Bourget J. M. et al. printed HUVECs and BMSCs together by
17
18 laser-assisted bioprinting to avoid HUVECs spreading out of the printed collagen structure 148.
19
20
21 What’s more, deposition of multiple cells in arranged postion can facilitate construction of
22
23 complex tissues. For example, co-printing of four bioinks including poly(dimethyl siloxane),
24
25
26 Pluronic F127 (PF127), GelMA encapsulating human neonatal dermal fibroblasts (hNDF),
27
28 and GelMA encapsulating 10T/12 fibroblasts, is developed by Kolesky D. B. et al. to
29
30
fabricate vascularized, heterogeneous tissue constructs 149 (Figure 4D). Besides, paracrine
31
32
33 signaling can be regulated by changing cell status, subsequently affecting cell functions. For
34
35 example, Snyder J. et al. reported that stem cells with ruptured membrane caused by elastic
36
37
38 and inelastic strain in bioprinting will release necrotic distress signals, which could initiate
39
40 migratory, biochemical synthesis and differentiation behavior in MSCs 119. In addition, control
41
42
43 of cell density and viability impacts the local concentration of bioactive proteins, further
44
45 influencing stem cell differentiation.
46
47
48 4.2.3. Additional factors
49
50 Additional factors involved in bioengineering studies include cytokines, hormones, and
51
52
enzymes. Adding cocktail of factors for stem cell induction is considered as a proven
53
54
55 technique in 2D stem cell differentiation, which has been widely utilized in 3D differentiation
56
57
28
58
59
1
2
3
4 as well. One method is to supplement these chemicals to culture media immersing the printed
5
6 structure, similar to 2D differentiation. However, bioinks may hinder the transfer of additional
7
8
factors, as the cells are encapsulated in bioinks but not exposed directly to the solution.
9
10
11 Sometimes, this feature may lead to a worse outcome compared with 2D differentiation 150.
12
13 Hence, another method is proposed to immobilize the additional factors in bioinks, where the
14
15
16 better dispersion of these factors and interactions with stem cells can be achieved 151,152
17
18 (Figure 4E). Besides, Poldervaart M. T. et al. explored improved osteogenesis differentiation
19
20
21 of goat multipotent stromal cells by incorporation of bone morphogenetic protein-2 (BMP-2)
22
23 into bioinks 153. Since multi-factors can be printed into different places precisely with
24
25
26 pre-designed patterns, complex tissues can be achieved by regulating growth and
27
28 differentiation of cells in different position with different factors. Ker E. D. et al. bioprinted
29
30
BMP-2 and fibroblast growth factor-2 (FGF-2) with spatial difference to induce C2C12
31
32
33 myoblasts to osteoblasts and tendon cells respectively to generate musculoskeletal models 154.
34
35 Apart from directing differentiation, additional factors show other functions, such as
36
37
38 improvement of cell adhesion, migration and proliferation 126. Moreover, adding
39
40 biodegradable units in bioinks will allow adjustable degradation rate of materials to match
41
42
43 better with ECM production 155. West J. L. et al. found that incorporation of enzymatically
44
45 degradable oligopeptide sequences in PEG can modulate degradation rate since local enzymes
46
47
48 will recognize and function at enzymatic cleavage sites within the bioink, resulting in the
49
50 optimization of cell migration and tissue formation 156.
51
52
53
54
55 4.3. Mass transfer
56
57
29
58
59
1
2
3
4 4.3.1. Porosity
5
6 Bioinks show the porous property as the basic approach of mass transfer inside the construct.
7
8
It is involved in the essential requirement of biocompatibility to provide space for cell
9
10
11 spreading and migration, and enable the exchange of chemicals to facilitate cell growth 125,157
12
13 (Figure 4F). As for stem cells, factors dissolved in culture media need to be transferred to
14
15
16 induce differentiation, So the number, shape and size of micropores, makes a difference in the
17
18 efficiency of stem cell proliferation and differentiation 150,158. It was reported by Domingos M.
19
20
21 et al. that MSC attachment and migration was associated with the pore size and shape of
22
23 bioprinted PCL scaffold. These feature also affected the subsequent vascularization for further
24
25
26 mass transfer 159. It should be mentioned that only open pores can facilitate the cell function,
27
28 as they enable exchange of various chemicals inside and outside the bioinks, and access of
29
30
encapsulated cells to the implant are feasible through these pores or channels 160.
31
32
33
34
35 But on the other hand there is a negative effect of porosity on structure stability, which can be
36
37
38 aggravated when inner non-connected molecules are eluted off. For example, Duarte Campos
39
40 D. F. et al. found that agarose-chitosan blends, which has better porosity for cell migration
41
42
43 were unprintable due to unsatisfactory stability, unlike agarose and agarose-collagen
44
45 hydrogels which are widely used in bioprinting 135.
46
47
48
49
50 4.3.2. Vascularization
51
52
Porous bioinks ensure the mass transfer in the early stage when cells are evenly distributed in
53
54
55 the structure. However, with the degradation of bioinks and the maturation of printed tissue,
56
57
30
58
59
1
2
3
4 close-packed cells would impede the penetration of solution. It is reported that solid tissues
5
6 with the thickness over 200 µm have limited clinical application 161 and cells can hardly
7
8
survive in tissues with thickness over 1mm 162, which can be more serious for highly
9
10
11 vascularized tissues such as liver, kidney, spleen, etc 163. Vascularization has become one of
12
13 the bottleneck problems in the application of engineered 3D tissues with clinical-relevant
14
15
16 dimension 164.
17
18
19
20
21 With the help of 3D bioprinting technology, angiogenesis elements can be constructed with
22
23 desired patterns to facilitate vascularization 165 (Figure 4G). Co-printing vessels within
24
25
26 tissues are mostly employed for the construction of vessels with large diameter (>6mm) like
27
28 aortas. Direct bioprinting of fine vessel structure is inefficient under current techniques with
29
30
inadequate resolution, especially for capillaries, hindering fabrication of mature vascularized
31
32
33 tissues 166. Considering the complex structure of nature vessels, temporary supporting
34
35 materials is printed and then removed to assist the formation of hollow channels. In addition,
36
37
38 bioprinting of aggregates facilitates the construction of vessels of large scale 92.
39
40
41
42
43 Autogenous vascularization methodology focuses more on the potentials of stem cells rather
44
45 than bioprinting process. Stem cells with differentiation potential into vascular tissues,
46
47
48 including endothelial progenitor cells (EPC) 167 and MSC 168, have been utilized in the
49
50 bioprinting vascularized structure under proper induction. But the progress of either
51
52
vasculogenesis or angiogenesis is time-consuming, where the viability of complex organs
53
54
55 may decrease sharply before the completion of this progress 16. Therefore, Mironov V. et al.
56
57
31
58
59
1
2
3
4 proposed a hybrid method by co-printing of cell aggregates for fabrication of branching large
5
6 vessels and pre-vascularized tissue building blocks as a microvascular network, to form a
7
8
vascular network with different sizes and types of vessels for organ bioprinting 97. Moreover,
9
10
11 attention should be paid to the connection between printed vascular channels and host
12
13 vasculature 166.
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49 Figure 4. Microenvironmental factors modulate stem cell functions in long-term culture. (A) Bioink
50
51 stiffness affects osteogenic differentiation of MSC, stained by oil red O. Reproduced with permission
52
53
54 from ref 129. Copyright 2015 PloS one. (B) Fabricate surface nano-features to regulate cell
55
56 morphology and migration. Reproduced with permission from ref 130. Copyright 2016 Colloids and
57
32
58
59
1
2
3
Surfaces B: Biointerfaces. (C) Bioprinting hybrid bioactive structure of ECM to improve
4
5
6 biocompatibility and PCL as the framework. Reproduced with permission from ref 137. Copyright
7
8
2014 Nature Communications. (D) Co-printing of 10T1/2 fibroblasts (blue), hNDFs (green), HUVECs
9
10
11 (red) can facilitate the formation of heterogeneous tissue constructs by paracrine. Reproduced with
12
13 permission from ref 149. Copyright 2014 Advanced Materials. (E) Tethering RGD ligands onto
14
15
16 ɑ-cyclodextrin-modified PEG can enhance spreading of MSCs. Reproduced with permission from ref
17
18 152. Copyright 2016 Advanced Materials. (F) Porous internal structure of alginate/gelatin hydrogel is
19
20
21 essential for mass transfer and cell migration. Reproduced with permission from ref 125. Copyright
22
23 2016 PloS one. (G) HUVECs (red) printed in hNDF-laden (green) matrix form a linear vessel with the
24
25
lumen to support perfusion for tissue mature in long-term culture. Reproduced with permission from
26
27
28 ref 165. Copyright 2016 Proceedings of the National Academy of Sciences.
29
30
31
32
33 5. Bioinks regulating printability and stem cell functions
34
35 Bioinks are considered a vital element in stem cell bioprinting technology, since they function
36
37
38 as one of the bioprinting elements, delivery vehicle of stem cells and also a very important
39
40 aspect of microenvironment for subsequent culture. In this section, we elaborate the
41
42
43 requirements and rationale of bioink properties from the perspective of bioprinting process
44
45 and stem cells, respectively. Some literature discussing bioink materials can be refereed to for
46
47
48 more details 169,170.
49
50
51
52
5.1. Printability: requirements of bioprinting process
53
54
55 The status of bioinks can be briefly divided into three stages throughout the whole bioprinting
56
57
33
58
59
1
2
3
4 process: 1) before bioprinting: liquid/sol state ; 2) during bioprinting: rapid solidification and
5
6 3) after bioprinting: solid/gel state. A printable bioink ought to display different properties in
7
8
accordance with the demand of different stage.
9
10
11
12
13 Flowability is essential to bioinks in the first stage, not only for encapsulating stem cells
14
15
16 expediently but for providing more options of bioprinting parameters as well, like printing
17
18 speed. As higher viscosity and higher printing speed can both generate greater mechanical
19
20
21 forces that pose harm to stem cells, viscous bioinks limit the printing speed to maintain cell
22
23 survival 65,123
. In addition, different enabling bioprinting techniques show the distinguished
24
25
26 optimal range of bioink viscosity, where most bioinks with viscosity over 300 mPa·s can
27
28 merely be applied in microextrusion-based bioprinting 10.
29
30
31
32
33 In the second stage, to establish a pre-designed structure with accuracy, the ability of rapid
34
35 solidification under the mild condition is prerequisite for bioink properties, which is realized
36
37
38 by crosslinking in most cases. Ideally, crosslinking of bioinks is supposed to take place upon
39
40 reaching the substrate and no sooner than extrusion from printing head (inkjet and
41
42 25
43 microextrusion-based bioprinting) or printing ribbon (laser-assisted bioprinting) .
44
45 Meanwhile, the crosslinking condition should cause minimal cell injury or death. Generally,
46
47
48 crosslinking methods can be divided into physical crosslinking, chemical crosslinking and
49
50 enzymatic crosslinking. Physical crosslinking does not involve the formation of covalent
51
52
bonds and is reversible for most bioinks. Its mechanism includes ionic reaction, complexation
53
54
55 reaction and hydrogen bonding reaction, which often occurs under the variation of charges,
56
57
34
58
59
1
2
3
4 temperature, and pH-value. By polymerization, chemical crosslinking is irreversible and can
5
6 create more stable structures under covalent reaction. As one of the most significant methods
7
8
in chemical crosslinking, photo-crosslinking has been widely employed and is the main
9
10
11 approach for stereolithography-based bioprinting. Under irradiation, photo-initiators in
12
13 bioinks can generate excited molecules to activate polymerization of monomers and
14
15
68
16 oligomers . Enzymatic crosslinking happens when corresponding protease and suitable
17
18 conditions are provided. For instance, fibrinogen can be converted by thrombin into fibrin,
19
20
21 which is a typical process in blood coagulation 39.
22
23
24
25
26 In the last stage, printed structures are required to maintain the shape through the subsequent
27
28 cell culture period. Crosslinked bioinks should provide long-term stability in the culture
29
30
medium, while sustaining stem cells proliferation, migration, metabolism and morphogenesis.
31
32
33 In addition, swelling and/or shrinking of bioinks may influence the integrity, size and
34
35 resolution of printed structures 123
. These factors must be taken into account in the design of
36
37
38 printing process. However, the addition of stem cell in the bioinks may also affect fluid and
39
40 crosslinking properties. Therefore, a simple test of bioink physical properties without cells is
41
42
43 inadequate, and multiple experiments are necessary to optimize and standardize the bioink
44
45 properties and printing process.
46
47
48
49
50 5.2. Biocompatibility: requirements of stem cells
51
52
During bioprinting, stem cells are evenly distributed in bioinks. After bioprinting, crosslinked
53
54
55 bioinks provide a long-term microenvironment for stem cell culture. Therefore, interactions
56
57
35
58
59
1
2
3
4 between stem cells and extracellular bioinks have a significant influence on stem cell viability,
5
6 attachment 15, proliferation 135, differentiation 21, migration 126 and morphogenesis 171.
7
8
9
10
11 Stem cells can be harmed by mechanical stress during bioprinting process, restricting the use
12
13 of viscous bioinks 22. Meanwhile, the toxicity of all the chemicals involved in bioprinting and
14
15
16 subsequent culture should be checked to avoid cell death, including bioinks, additions like
17
18 photo initiators 172, by-products during crosslinking 123, and degradation products 173.
19
20
21
22
23 Stem cell attachment, migration and morphology are mainly impacted in post-printing culture.
24
25
26 Ligands between stem cells and polymer chains determine whether cells can attach to the
27
28 surface of bioinks facilely or detach for migration 159. What’s more, bioinks with high strength
29
30
and stiffness are disadvantageous for cell mobility. Thus chemical crosslinked bioinks usually
31
32
33 demonstrate less biocompatibility than physical crosslinked ones since they have higher
34
35 stability 152
. A common method to tune cell attachment, migration and formation of specific
36
37
38 morphology such as cell aggregations without changing the matrix of structure is surface
39
40 modification, including generation of nano-scale topographies 131
or bonding of chemical
41
42
43 ligands 152.
44
45
46
47
48 Provided with 3D microenvironment, stem cells exhibit better proliferation in printed
49
50 structure compared with 2D culture 144. Increased proliferation can be achieved by optimizing
51
52 135
alignment of bioinks for some stem cells like MSCs . Porosity is another factor regulating
53
54
126
55 stem cell proliferation by controlling the transfer of oxygen, nutrition and metabolite . In
56
57
36
58
59
1
2
3
4 addition, the degradation rate of bioinks should be in accordance with proliferation rate of
5
6 stem cells and production of stem cell ECM, in order to offer adequate space for cell growth
7
8
and tissue maturation 15.
9
10
11
12
13 Distinct from bioprinting of primary cells, differentiation is one of the most important feature
14
15
16 of stem cells. On the one hand, bioinks should not result in undesired differentiation, which
17
18 may occur in dECM and some other natural bioinks. On the other hand, stem cell
19
20
21 differentiation can be modulated by change of physical properties or chemical components.
22
23 Thus, a comprehensive optimization of bioinks is valuable for improved stem cell
24
25
26 differentiation.
27
28
29
30
In all, the choice of bioinks involves a delicate balance between printability and
31
32
33 biocompatibility. Generally, natural bioinks offers better support for stem cell functions, but
34
35 usually shows poorer mechanical properties and repeatability, probably due to batch-to-batch
36
37
38 variations. Artificial bioinks are more stable and controllable, but demonstrate less
39
40 biocompatible in most cases. Most commonly used bioinks for stem cell bioprinting are listed
41
42
43 in Table 2.
44
45
46
47
48
49
50
51
52
53
54
55
56
57
37
58
59
1
2
3
4
5
6
7
8
9
10
11 Table 2. Commonly used bioinks in stem cell bioprinting
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
38
58
59
1
2
3
Bioprinting technique Crosslinking method Cell type Viability(%) Ref.
4
5 Agarose Micrextrusion-based Thermal MSCs 98.8 135
6 bioprinting
7 Agarose/alginate/chitosan Micrextrusion-based Ionically NSCs >75 62

8 bioprinting
9 Alginate Micrextrusion-based Ionically MSCs 95 46
10 bioprinting
11 Inkjet bioprinting Ionically ESCs, iPSCs 90 21
12
13 Alginate/Collagen type I Micrextrusion-based Ionically ASCs 91 54
14 bioprinting
15 Micrextrusion-based Ionically ASCs, preosteoblasts 92.3 174
16 bioprinting
17
Laser-assisted bioprinting Ionically MSCs 100 71
18
19
Alginate/gelatin Micrextrusion-based Thermal, ionically ESCs 90 27
20 bioprinting
21
22 Micrextrusion-based Ionically Glioma stem cells 86.9 63
bioprinting
23
24 Micrextrusion-based Ionically MSCs 89 125
bioprinting
25
26 Chitosan Micrextrusion-based pH mediated ASCs - 175
bioprinting
27
28 Collagen type I Inkjet bioprinting pH mediated MSCs 81 102
29
30 Collagen type I/fibrin Inkjet bioprinting pH mediated, enzymatic NSCs 92.9 19
31
32 Laser-assisted bioprinting Enzymatic Amniotic fluid-derived - 104
33 stem cells
34 Fibrin Inkjet bioprinting Enzymatic MSCs - 102
35
36 Gelatin inkjet bioprinting Thermal, polymerized MSCs - 102
37
38 Laser-assisted bioprinting Thermal ESCs - 73
39
40
GelMa Inkjet bioprinting Photopolymerized MSCs 90 18
41
42
Stereolithography-based Photopolymerized BMSCs - 77
43 bioprinting
44
Stereolithography-based Photopolymerized NSCs - 82
45 bioprinting
46
47 GelMa/PEG Inkjet bioprinting Photopolymerized MSCs >80 39
48
49 Hyaluronic acid/matrigel Micrextrusion-based Polymerized MSCs - 176
bioprinting
50
51 PEG Micrextrusion-based Photopolymerized MSCs 90 177
bioprinting
52
53 polyurethane Micrextrusion-based Thermal NSCs >50 61
54 bioprinting
55
56 6. Perspective
57
39
58
59
1
2
3
4 Stem cell bioprinting takes advantage of both the precise positioning of the substance of
5
6 bioprinting technology and the proliferation and differentiation abilities of stem cells,
7
8
showing great potential in stem cell biology, drug testing, tissue engineering and regenerative
9
10
11 medicine research. But this emerging technology faces many challenges related to cellular,
12
13 technical, and material aspects in the whole lifespan. In recent years, significant progress has
14
15
16 been achieved in improving stem cell functions and creating more complex structures.
17
18
19
20
21 Creation of microenvironment comparable to natural tissue/organ is proposed as a direction to
22
23 improve stem cell behaviors in printed structure. On the one hand, the invention of novel
24
25
26 printable biomimetic materials is of significant importance. Composite materials can be
27
28 utilized as bioinks combining the printability of synthetic materials and biocompatibility of
29
30
natural materials. Besides, functional bioinks can be also generated by extra additions or
31
32
33 surface modification, which ensure better proliferation, induced differentiation and intelligent
34
35 degradation 173
. On the other hand, precise spatial distribution of multiple bioinks with
36
37
38 suitable mechanical properties and other chemicals in accordance with the stem cell demands
39
40 can help regulate microenvironment in micro-scale. Merceron T. K. et al. reported that a
41
42
43 muscle-tendon unit construct was fabricated by bioprinting four bioinks encapsulated with
44
45 myoblasts and fibroblasts with regionally defined characteristics 178
. In addition, to predict
46
47
48 cell behaviors in the microenvironment, computer modeling has been utilized in the research
49
50 of post-printing cell culture, including kinetic Monte Carlo (KMC) simulations 179
, cellular
51
52 180
particle dynamics (CPD) and phase field theories 96. Currently, these modeling techniques
53
54
55 are mainly focused on cell aggregate fusion or stress distribution inside the structures.
56
57
40
58
59
1
2
3
4 Simulation of flow velocity, stress, and temperature distribution in the fluid to predict stem
5
6 cell status in bioinks during bioprinting is necessary. Further development in the modeling of
7
8
biochemical environment around stem cells after bioprinting is more desired. In the future,
9
10
11 modeling of stem cell situation in bioinks from loading to long-term culture is needed.
12
13
14
15
16 Another significant direction is scale-up and mass production for clinical and/or commercial
17
18 application. On one hand, high throughput technology is prerequisite for mass production
19
20
21 with quality controllability. On the other hand, commercial availability and practicability of
22
23 stem cells and bioinks should be also considered, including source, cost, shelf life and FDA
24
25
26 administration. In addition, equipment and technologies should be appropriate for scale
27
28 production. Before bioprinting, stem cells are supposed to proliferate in controlled ways to
29
30
provide adequate cell numbers for bioprinting. Pre-differentiation of stem cells and
31
32
33 pre-fabrication of cell aggregates in quantity are sometimes required in specific research.
34
35 After bioprinting, structures with stem cells are proposed to be cultured in bioreactors for
36
37
38 fusion of cells and maturation of tissues. A further step is the establishment of assembly line
39
40 for organ biofabrication, whose components involve clinical cell sorters, stem cell
41
42
43 propagation bioreactors, cell differentiators, cell aggregate biofabricators, cell encapsulators,
44
45 robotic bioprinters, and perfusion bioreactors. The whole assembly line should be automatic,
46
47 181
48 biocompatible, large-scale, and flexible for generation of diverse tissues and organs . A
49
50 real-time monitor of cell status is suggested to inform researchers and regulate fabrication
51
52
process. What’s more, since most laboratories around the world utilize home-made or
53
54
55 customized 3D bioprinters for specific research nowadays, standardization is another critical
56
57
41
58
59
1
2
3 182
4 step in the industrialization of stem cell bioprinting . Tissues and organs should be
5
6 generated by the standardized process in a reproducible way.
7
8
9
10
11 In the aspect of clinical application, stem cell bioprinting shows great potential in precision
12
13 medicine. Stem cells are obtained directly from a patient or modified with individual genetic
14
15
16 content. By bioprinting and subsequent differentiation, specific tissues or organs are available
17
18 as either in vitro testing models or in vivo implants for diagnosis, analysis, and treatment of
19
20
21 personal disease. In recent years, bioprinting of iPSCs offers a potential versatile approach for
22
23 production of customized tissues, as this type of cells can be acquired from every patient by
24
25
26 reprogramming of adult cells, and differentiate into desired cell types 21. Though challenges
27
28 still exist in the functional stability and security of iPSCs and impact of bioprinting process on
29
30
iPSCs, but the application of iPSC bioprinting hold a bright future for precision medicine.
31
32
33
34
35 In conclusion, even still in early stage, 3D bioprinting of stem cells is emerging as a
36
37
38 promising technology which has potential applications in many research. Bioprinting
39
40 technology provides a convenient and repeatable approach for precise spatial positioning of
41
42
43 both cells and bioink, which would facilitate the delicate study of cell niche regulation, which
44
45 is hardly feasible in other 3D culture methods. Meanwhile, stem cells serve as the reliable cell
46
47
48 sources in bioprinting for tissue engineering, regenerative medicine, drug testing and cancer
49
50 studies. Their proliferation and self-renewal property facilitates rapid fabrication of
51
52 183
large-scale tissues or even organs on demand such as healing of large skin wounds . In
53
54
55 addition, induced differentiation of stem cells in bioprinted structure can be combined with
56
57
42
58
59
1
2
3
4 organoid technology and work as a versatile tool for construction of complex functional
5
6 tissues of various types. With enhanced understanding of the interplay between stem cells and
7
8
microenvironment and improvement of bioprinting process, bioinks and stem cell technology,
9
10
11 future application of 3D bioprinting of stem cells can be extended to drug development, organ
12
13 transplantation and personalized medicine as a reliable, convenient and efficient technology.
14
15
16
17
18
19
20
21
22
23 Acknowledgements
24
25
26 The authors acknowledge fundings from the National Natural Science Foundation of China
27
28 (No. 31500818 and No. 31711530218).
29
30
31
32
33
34
35
36
37
38 References
39
40 [1] Beltrami, A. P.; Barlucchi, L.; Torella, D.; Baker, M.; Limana, F.; Chimenti, S.; Kasahara, H.; Rota,
41
42
43 M.; Musso, E.; Urbanek, K.; Leri, A.; Kajstura, J.; Nadal-Ginard, B.; Anversa, P., Adult cardiac stem
44
45 cells are multipotent and support myocardial regeneration. Cell 2003, 114 (6), 763-76.
46
47
[2] Thomson, J. A.; Itskovitz-Eldor, J.; Shapiro, S. S.; Waknitz, M. A.; Swiergiel, J. J.; Marshall, V. S.;
48
49
50 Jones, J. M., Embryonic stem cell lines derived from human blastocysts. Science 1998, 282 (5391),
51
52
1145-7.
53
54
55 [3] Tran, C.; Damaser, M. S., Stem cells as drug delivery methods: application of stem cell secretome
56
57
43
58
59
1
2
3
for regeneration. Advanced drug delivery reviews 2015, 82-83, 1-11. DOI: 10.1016/j.addr.2014.10.007.
4
5
6 [4] Gage, F. H., Mammalian neural stem cells. Science 2000, 287 (5457), 1433-8.
7
8
[5] Reya, T.; Morrison, S. J.; Clarke, M. F.; Weissman, I. L., Stem cells, cancer, and cancer stem cells.
9
10
11 Nature 2001, 414 (6859), 105-11. DOI: 10.1038/35102167.
12
13 [6] Pittenger, M. F.; Mackay, A. M.; Beck, S. C.; Jaiswal, R. K.; Douglas, R.; Mosca, J. D.; Moorman,
14
15
16 M. A.; Simonetti, D. W.; Craig, S.; Marshak, D. R., Multilineage potential of adult human
17
18 mesenchymal stem cells. Science 1999, 284 (5411), 143-7.
19
20
21 [7] Zuk, P. A.; Zhu, M.; Ashjian, P.; De Ugarte, D. A.; Huang, J. I.; Mizuno, H.; Alfonso, Z. C.; Fraser,
22
23 J. K.; Benhaim, P.; Hedrick, M. H., Human adipose tissue is a source of multipotent stem cells.
24
25
Molecular biology of the cell 2002, 13 (12), 4279-95. DOI: 10.1091/mbc.e02-02-0105.
26
27
28 [8] Takahashi, K.; Yamanaka, S., Induction of pluripotent stem cells from mouse embryonic and adult
29
30
fibroblast cultures by defined factors. Cell 2006, 126 (4), 663-76. DOI: 10.1016/j.cell.2006.07.024.
31
32
33 [9] Chua, C.K.; Yeong, W. Y., Bioprinting: principles and applications. World Scientific Publishing Co
34
35 Inc 2014.
36
37
38 [10] Doyle, K., Bioprinting: from patches to parts. Genetic Engineering & Biotechnology News 2014,
39
40 34(10): 1, 34-35.
41
42
43 [11] Hospodiuk, M.; Dey, M.; Sosnoski, D.; Ozbolat, I. T., The bioink: A comprehensive review on
44
45 bioprintable materials. Biotechnol Adv 2017, 35 (2), 217-239. DOI: 10.1016/j.biotechadv.2016.12.006.
46
47
[12] Mehesz, A. N.; Brown, J.; Hajdu, Z.; Beaver, W.; da Silva, J. V.; Visconti, R. P.; Markwald, R. R.;
48
49
50 Mironov, V., Scalable robotic biofabrication of tissue spheroids. Biofabrication 2011, 3 (2), 025002.
51
52
DOI: 10.1088/1758-5082/3/2/025002.
53
54
55 [13] Goral, V. N.; Au, S. H.; Faris, R. A.; Yuen, P. K., Microstructured multi-well plate for
56
57
44
58
59
1
2
3
three-dimensional packed cell seeding and hepatocyte cell culture. Biomicrofluidics 2014, 8 (4),
4
5
6 046502. DOI: 10.1063/1.4892978.
7
8
[14] Tasoglu, S.; Demirci, U., Bioprinting for stem cell research. Trends in biotechnology 2013, 31 (1),
9
10
11 10-9. DOI: 10.1016/j.tibtech.2012.10.005.
12
13 [15] Murphy, S. V.; Atala, A., 3D bioprinting of tissues and organs. Nature biotechnology 2014, 32 (8),
14
15
16 773-85. DOI: 10.1038/nbt.2958.
17
18 [16] Guillotin, B.; Guillemot, F., Cell patterning technologies for organotypic tissue fabrication. Trends
19
20
21 in biotechnology 2011, 29 (4), 183-90. DOI: 10.1016/j.tibtech.2010.12.008.
22
23 [17] Nakamura, M.; Iwanaga, S.; Henmi, C.; Arai, K.; Nishiyama, Y., Biomatrices and biomaterials for
24
25
future developments of bioprinting and biofabrication. Biofabrication 2010, 2 (1), 014110. DOI:
26
27
28 10.1088/1758-5082/2/1/014110.
29
30
[18] Gurkan, U. A.; El Assal, R.; Yildiz, S. E.; Sung, Y.; Trachtenberg, A. J.; Kuo, W. P.; Demirci, U.,
31
32
33 Engineering anisotropic biomimetic fibrocartilage microenvironment by bioprinting mesenchymal stem
34
35 cells in nanoliter gel droplets. Molecular pharmaceutics 2014, 11 (7), 2151-9. DOI:
36
37
38 10.1021/mp400573g.
39
40 [19] Lee, Y. B.; Polio, S.; Lee, W.; Dai, G.; Menon, L.; Carroll, R. S.; Yoo, S. S., Bio-printing of
41
42
43 collagen and VEGF-releasing fibrin gel scaffolds for neural stem cell culture. Experimental neurology
44
45 2010, 223 (2), 645-52. DOI: 10.1016/j.expneurol.2010.02.014.
46
47
[20] Obregon, F.; Vaquette, C.; Ivanovski, S.; Hutmacher, D. W.; Bertassoni, L. E., Three-Dimensional
48
49
50 Bioprinting for Regenerative Dentistry and Craniofacial Tissue Engineering. Journal of dental research
51
52
2015, 94 (9 Suppl), 143S-52S. DOI: 10.1177/0022034515588885.
53
54
55 [21] Faulkner-Jones, A.; Fyfe, C.; Cornelissen, D. J.; Gardner, J.; King, J.; Courtney, A.; Shu, W.,
56
57
45
58
59
1
2
3
Bioprinting of human pluripotent stem cells and their directed differentiation into hepatocyte-like cells
4
5
6 for the generation of mini-livers in 3D. Biofabrication 2015, 7 (4), 044102. DOI:
7
8
10.1088/1758-5090/7/4/044102.
9
10
11 [22] Malda, J.; Visser, J.; Melchels, F. P.; Jungst, T.; Hennink, W. E.; Dhert, W. J.; Groll, J.; Hutmacher,
12
13 D. W., 25th anniversary article: Engineering hydrogels for biofabrication. Advanced materials 2013, 25
14
15
16 (36), 5011-28. DOI: 10.1002/adma.201302042.
17
18 [23] Muller, M.; Becher, J.; Schnabelrauch, M.; Zenobi-Wong, M., Nanostructured Pluronic hydrogels
19
20
21 as bioinks for 3D bioprinting. Biofabrication 2015, 7 (3), 035006. DOI:
22
23 10.1088/1758-5090/7/3/035006.
24
25
[24] Pek, Y. S.; Wan, A. C.; Ying, J. Y., The effect of matrix stiffness on mesenchymal stem cell
26
27
28 differentiation in a 3D thixotropic gel. Biomaterials 2010, 31 (3), 385-91. DOI:
29
30
10.1016/j.biomaterials.2009.09.057.
31
32
33 [25] McNamara, L. E.; McMurray, R. J.; Biggs, M. J.; Kantawong, F.; Oreffo, R. O.; Dalby, M. J.,
34
35 Nanotopographical control of stem cell differentiation. Journal of tissue engineering 2010, 2010,
36
37
38 120623. DOI: 10.4061/2010/120623.
39
40 [26] Pati, F.; Jang, J.; Ha, D. H.; Won Kim, S.; Rhie, J. W.; Shim, J. H.; Kim, D. H.; Cho, D. W.,
41
42
43 Printing three-dimensional tissue analogues with decellularized extracellular matrix bioink. Nature
44
45 communications 2014, 5, 3935. DOI: 10.1038/ncomms4935.
46
47
[27] Ouyang, L.; Yao, R.; Zhao, Y.; Sun, W., Effect of bioink properties on printability and cell viability
48
49
50 for 3D bioplotting of embryonic stem cells. Biofabrication 2016, 8 (3), 035020. DOI:
51
52
10.1088/1758-5090/8/3/035020.
53
54
55 [28] Zhao, Y.; Li, Y.; Mao, S.; Sun, W.; Yao, R., The influence of printing parameters on cell survival
56
57
46
58
59
1
2
3
rate and printability in microextrusion-based 3D cell printing technology. Biofabrication 2015, 7 (4),
4
5
6 045002. DOI: 10.1088/1758-5090/7/4/045002.
7
8
[29] Gao, G.; Cui, X., Three-dimensional bioprinting in tissue engineering and regenerative medicine.
9
10
11 Biotechnology letters 2016, 38 (2), 203-11. DOI: 10.1007/s10529-015-1975-1.
12
13 [30] Norotte, C.; Marga, F. S.; Niklason, L. E.; Forgacs, G., Scaffold-free vascular tissue engineering
14
15
16 using bioprinting. Biomaterials 2009, 30 (30), 5910-7. DOI: 10.1016/j.biomaterials.2009.06.034.
17
18 [31] Yamaguchi, S.; Ueno, A.; Akiyama, Y.; Morishima, K., Cell patterning through inkjet printing of
19
20
21 one cell per droplet. Biofabrication 2012, 4 (4), 045005. DOI: 10.1088/1758-5082/4/4/045005.
22
23 [32] Roth, E. A.; Xu, T.; Das, M.; Gregory, C.; Hickman, J. J.; Boland, T., Inkjet printing for
24
25
high-throughput cell patterning. Biomaterials 2004, 25 (17), 3707-15. DOI:
26
27
28 10.1016/j.biomaterials.2003.10.052.
29
30
[33] Calvert, P., Materials science. Printing cells. Science 2007, 318 (5848), 208-9. DOI:
31
32
33 10.1126/science.1144212.
34
35 [34] Cui, X.; Gao, G.; Yonezawa, T.; Dai, G., Human cartilage tissue fabrication using
36
37
38 three-dimensional inkjet printing technology. Journal of visualized experiments : JoVE 2014, (88).
39
40 DOI: 10.3791/51294.
41
42
43 [35] Lorber, B.; Hsiao, W. K.; Hutchings, I. M.; Martin, K. R., Adult rat retinal ganglion cells and glia
44
45 can be printed by piezoelectric inkjet printing. Biofabrication 2014, 6 (1), 015001. DOI:
46
47
10.1088/1758-5082/6/1/015001.
48
49
50 [36] Faulkner-Jones, A.; Greenhough, S.; King, J. A.; Gardner, J.; Courtney, A.; Shu, W., Development
51
52
of a valve-based cell printer for the formation of human embryonic stem cell spheroid aggregates.
53
54
55 Biofabrication 2013, 5 (1), 015013. DOI: 10.1088/1758-5082/5/1/015013.
56
57
47
58
59
1
2
3
[37] Yamada, M.; Imaishi, H.; Morigaki, K., Microarrays of phospholipid bilayers generated by inkjet
4
5
6 printing. Langmuir : the ACS journal of surfaces and colloids 2013, 29 (21), 6404-8. DOI:
7
8
10.1021/la400570h.
9
10
11 [38] Yao, R.; Zhang, R.; Lin, F.; Luan, J., Injectable cell/hydrogel microspheres induce the formation of
12
13 fat lobule-like microtissues and vascularized adipose tissue regeneration. Biofabrication 2012, 4 (4),
14
15
16 045003. DOI: 10.1088/1758-5082/4/4/045003.
17
18 [39] Gao, G.; Schilling, A. F.; Hubbell, K.; Yonezawa, T.; Truong, D.; Hong, Y.; Dai, G.; Cui, X.,
19
20
21 Improved properties of bone and cartilage tissue from 3D inkjet-bioprinted human mesenchymal stem
22
23 cells by simultaneous deposition and photocrosslinking in PEG-GelMA. Biotechnology letters 2015, 37
24
25
(11), 2349-55. DOI: 10.1007/s10529-015-1921-2.
26
27
28 [40] Gao, G.; Hubbell, K.; Schilling, A. F.; Dai, G.; Cui, X., Bioprinting Cartilage Tissue from
29
30
Mesenchymal Stem Cells and PEG Hydrogel. Methods in molecular biology 2017, 1612, 391-398. DOI:
31
32
33 10.1007/978-1-4939-7021-6_28.
34
35 [41] Phillippi, J. A.; Miller, E.; Weiss, L.; Huard, J.; Waggoner, A.; Campbell, P., Microenvironments
36
37
38 engineered by inkjet bioprinting spatially direct adult stem cells toward muscle- and bone-like
39
40 subpopulations. Stem cells 2008, 26 (1), 127-34. DOI: 10.1634/stemcells.2007-0520.
41
42
43 [42] Ilkhanizadeh, S.; Teixeira, A. I.; Hermanson, O., Inkjet printing of macromolecules on hydrogels
44
45 to steer neural stem cell differentiation. Biomaterials 2007, 28 (27), 3936-43. DOI:
46
47
10.1016/j.biomaterials.2007.05.018.
48
49
50 [43] Xu, T.; Zhao, W.; Zhu, J. M.; Albanna, M. Z.; Yoo, J. J.; Atala, A., Complex heterogeneous tissue
51
52
constructs containing multiple cell types prepared by inkjet printing technology. Biomaterials 2013, 34
53
54
55 (1), 130-9. DOI: 10.1016/j.biomaterials.2012.09.035.
56
57
48
58
59
1
2
3
[44] Cui, X.; Dean, D.; Ruggeri, Z. M.; Boland, T., Cell damage evaluation of thermal inkjet printed
4
5
6 Chinese hamster ovary cells. Biotechnology and bioengineering 2010, 106 (6), 963-9. DOI:
7
8
10.1002/bit.22762.
9
10
11 [45] Xu, T.; Rohozinski, J.; Zhao, W.; Moorefield, E. C.; Atala, A.; Yoo, J. J., Inkjet-mediated gene
12
13 transfection into living cells combined with targeted delivery. Tissue engineering. Part A 2009, 15 (1),
14
15
16 95-101. DOI: 10.1089/ten.tea.2008.0095.
17
18 [46] Fedorovich, N. E.; De Wijn, J. R.; Verbout, A. J.; Alblas, J.; Dhert, W. J., Three-dimensional fiber
19
20
21 deposition of cell-laden, viable, patterned constructs for bone tissue printing. Tissue engineering. Part
22
23 A 2008, 14 (1), 127-33. DOI: 10.1089/ten.a.2007.0158.
24
25
[47] Jones, N., Science in three dimensions: the print revolution. Nature 2012, 487 (7405), 22-3. DOI:
26
27
28 10.1038/487022a.
29
30
[48] Wilson, S. A.; Cross, L. M.; Peak, C. W.; Gaharwar, A. K., Shear-Thinning and Thermo-Reversible
31
32
33 Nanoengineered Inks for 3D Bioprinting. ACS applied materials & interfaces 2017, 9 (50),
34
35 43449-43458. DOI: 10.1021/acsami.7b13602.
36
37
38 [49] Derby, B., Printing and prototyping of tissues and scaffolds. Science 2012, 338 (6109), 921-6. DOI:
39
40 10.1126/science.1226340.
41
42
43 [50] Ferris, C. J.; Gilmore, K. G.; Wallace, G. G.; In het Panhuis, M., Biofabrication: an overview of
44
45 the approaches used for printing of living cells. Applied microbiology and biotechnology 2013, 97 (10),
46
47
4243-58. DOI: 10.1007/s00253-013-4853-6.
48
49
50 [51] Reid, J. A.; Mollica, P. A.; Johnson, G. D.; Ogle, R. C.; Bruno, R. D.; Sachs, P. C., Accessible
51
52
bioprinting: adaptation of a low-cost 3D-printer for precise cell placement and stem cell differentiation.
53
54
55 Biofabrication 2016, 8 (2), 025017. DOI: 10.1088/1758-5090/8/2/025017.
56
57
49
58
59
1
2
3
[1] Costantini, M.; Testa, S.; Mozetic, P.; Barbetta, A.; Fuoco, C.; Fornetti, E.; Tamiro, F.; Bernardini,
4
5
6 S.; Jaroszewicz, J.; Swieszkowski, W.; Trombetta, M.; Castagnoli, L.; Seliktar, D.; Garstecki, P.;
7
8
Cesareni, G.; Cannata, S.; Rainer, A.; Gargioli, C., Microfluidic-enhanced 3D bioprinting of aligned
9
10
11 myoblast-laden hydrogels leads to functionally organized myofibers in vitro and in vivo. Biomaterials
12
13 2017, 131, 98-110. DOI: 10.1016/j.biomaterials.2017.03.026.
14
15
16 [53] Duchi, S.; Onofrillo, C.; O'Connell, C. D.; Blanchard, R.; Augustine, C.; Quigley, A. F.; Kapsa, R.
17
18 M. I.; Pivonka, P.; Wallace, G.; Di Bella, C.; Choong, P. F. M., Handheld Co-Axial Bioprinting:
19
20
21 Application to in situ surgical cartilage repair. Scientific reports 2017, 7 (1), 5837. DOI:
22
23 10.1038/s41598-017-05699-x.
24
25
[54] Yeo, M.; Lee, J. S.; Chun, W.; Kim, G. H., An Innovative Collagen-Based Cell-Printing Method
26
27
28 for Obtaining Human Adipose Stem Cell-Laden Structures Consisting of Core-Sheath Structures for
29
30
Tissue Engineering. Biomacromolecules 2016, 17 (4), 1365-75. DOI: 10.1021/acs.biomac.5b01764.
31
32
33 [55] Song, S. J.; Choi, J.; Park, Y. D.; Hong, S.; Lee, J. J.; Ahn, C. B.; Choi, H.; Sun, K., Sodium
34
35 alginate hydrogel-based bioprinting using a novel multinozzle bioprinting system. Artificial organs
36
37
38 2011, 35 (11), 1132-6. DOI: 10.1111/j.1525-1594.2011.01377.x.
39
40 [56] Ahn, S. H.; Lee, H. J.; Lee, J. S.; Yoon, H.; Chun, W.; Kim, G. H., A novel cell-printing method
41
42
43 and its application to hepatogenic differentiation of human adipose stem cell-embedded mesh structures.
44
45 Scientific reports 2015, 5, 13427. DOI: 10.1038/srep13427.
46
47
[57] Highley, C. B.; Rodell, C. B.; Burdick, J. A., Direct 3D Printing of Shear-Thinning Hydrogels into
48
49
50 Self-Healing Hydrogels. Advanced materials 2015, 27 (34), 5075-9. DOI: 10.1002/adma.201501234.
51
52
[58] Gao, G.; Schilling, A. F.; Yonezawa, T.; Wang, J.; Dai, G.; Cui, X., Bioactive nanoparticles
53
54
55 stimulate bone tissue formation in bioprinted three-dimensional scaffold and human mesenchymal stem
56
57
50
58
59
1
2
3
cells. Biotechnology journal 2014, 9 (10), 1304-11. DOI: 10.1002/biot.201400305.
4
5
6 [59] Yeo, M.; Ha, J.; Lee, H.; Kim, G., Fabrication of hASCs-laden structures using extrusion-based
7
8
cell printing supplemented with an electric field. Acta biomaterialia 2016, 38, 33-43. DOI:
9
10
11 10.1016/j.actbio.2016.04.017.
12
13 [60] Kang, L. H.; Armstrong, P. A.; Lee, L. J.; Duan, B.; Kang, K. H.; Butcher, J. T., Optimizing
14
15
16 Photo-Encapsulation Viability of Heart Valve Cell Types in 3D Printable Composite Hydrogels. Annals
17
18 of biomedical engineering 2017, 45 (2), 360-377. DOI: 10.1007/s10439-016-1619-1.
19
20
21 [61] Hsieh, F. Y.; Lin, H. H.; Hsu, S. H., 3D bioprinting of neural stem cell-laden thermoresponsive
22
23 biodegradable polyurethane hydrogel and potential in central nervous system repair. Biomaterials 2015,
24
25
71, 48-57. DOI: 10.1016/j.biomaterials.2015.08.028.
26
27
28 [62] Gu, Q.; Tomaskovic-Crook, E.; Lozano, R.; Chen, Y.; Kapsa, R. M.; Zhou, Q.; Wallace, G. G.;
29
30
Crook, J. M., Functional 3D Neural Mini-Tissues from Printed Gel-Based Bioink and Human Neural
31
32
33 Stem Cells. Advanced healthcare materials 2016, 5 (12), 1429-38. DOI: 10.1002/adhm.201600095.
34
35 [63] Dai, X.; Ma, C.; Lan, Q.; Xu, T., 3D bioprinted glioma stem cells for brain tumor model and
36
37
38 applications of drug susceptibility. Biofabrication 2016, 8 (4), 045005. DOI:
39
40 10.1088/1758-5090/8/4/045005.
41
42
43 [64] Gu, Q.; Tomaskovic-Crook, E.; Wallace, G. G.; Crook, J. M., 3D Bioprinting Human Induced
44
45 Pluripotent Stem Cell Constructs for In Situ Cell Proliferation and Successive Multilineage
46
47
Differentiation. Advanced healthcare materials 2017, 6 (17). DOI: 10.1002/adhm.201700175.
48
49
50 [65] Smith, C. M.; Stone, A. L.; Parkhill, R. L.; Stewart, R. L.; Simpkins, M. W.; Kachurin, A. M.;
51
52
Warren, W. L.; Williams, S. K., Three-dimensional bioassembly tool for generating viable
53
54
55 tissue-engineered constructs. Tissue engineering 2004, 10 (9-10), 1566-76. DOI:
56
57
51
58
59
1
2
3
10.1089/ten.2004.10.1566.
4
5
6 [66] Chang, R.; Nam, J.; Sun, W., Effects of dispensing pressure and nozzle diameter on cell survival
7
8
from solid freeform fabrication-based direct cell writing. Tissue engineering. Part A 2008, 14 (1), 41-8.
9
10
11 DOI: 10.1089/ten.a.2007.0004.
12
13 [67] Ringeisen, B. R.; Kim, H.; Barron, J. A.; Krizman, D. B.; Chrisey, D. B.; Jackman, S.; Auyeung, R.
14
15
16 Y.; Spargo, B. J., Laser printing of pluripotent embryonal carcinoma cells. Tissue engineering 2004, 10
17
18 (3-4), 483-91. DOI: 10.1089/107632704323061843.
19
20
21 [68] Boland, T.; Xu, T.; Damon, B.; Cui, X., Application of inkjet printing to tissue engineering.
22
23 Biotechnology journal 2006, 1 (9), 910-7. DOI: 10.1002/biot.200600081.
24
25
[69] Koch, L.; Gruene, M.; Unger, C.; Chichkov, B., Laser assisted cell printing. Current
26
27
28 pharmaceutical biotechnology 2013, 14 (1), 91-7.
29
30
[70] Guillotin, B.; Souquet, A.; Catros, S.; Duocastella, M.; Pippenger, B.; Bellance, S.; Bareille, R.;
31
32
33 Remy, M.; Bordenave, L.; Amedee, J.; Guillemot, F., Laser assisted bioprinting of engineered tissue
34
35 with high cell density and microscale organization. Biomaterials 2010, 31 (28), 7250-6. DOI:
36
37
38 10.1016/j.biomaterials.2010.05.055.
39
40 [71] Ali, M.; Pages, E.; Ducom, A.; Fontaine, A.; Guillemot, F., Controlling laser-induced jet formation
41
42
43 for bioprinting mesenchymal stem cells with high viability and high resolution. Biofabrication 2014, 6
44
45 (4), 045001. DOI: 10.1088/1758-5082/6/4/045001.
46
47
[72] Sorkio, A.; Koch, L.; Koivusalo, L.; Deiwick, A.; Miettinen, S.; Chichkov, B.; Skottman, H.,
48
49
50 Human stem cell based corneal tissue mimicking structures using laser-assisted 3D bioprinting and
51
52
functional bioinks. Biomaterials 2018, 171, 57-71. DOI: 10.1016/j.biomaterials.2018.04.034.
53
54
55 [73] Dias, A. D.; Unser, A. M.; Xie, Y.; Chrisey, D. B.; Corr, D. T., Generating size-controlled
56
57
52
58
59
1
2
3
embryoid bodies using laser direct-write. Biofabrication 2014, 6 (2), 025007. DOI:
4
5
6 10.1088/1758-5082/6/2/025007.
7
8
[74] Arcaute, K.; Mann, B. K.; Wicker, R. B., Stereolithography of three-dimensional bioactive
9
10
11 poly(ethylene glycol) constructs with encapsulated cells. Annals of biomedical engineering 2006, 34
12
13 (9), 1429-41. DOI: 10.1007/s10439-006-9156-y.
14
15
16 [75] Skoog, S. A.; Goering, P. L.; Narayan, R. J., Stereolithography in tissue engineering. Journal of
17
18 materials science. Materials in medicine 2014, 25 (3), 845-56. DOI: 10.1007/s10856-013-5107-y.
19
20
21 [76] Wang, Z.; Abdulla, R.; Parker, B.; Samanipour, R.; Ghosh, S.; Kim, K., A simple and
22
23 high-resolution stereolithography-based 3D bioprinting system using visible light crosslinkable bioinks.
24
25
Biofabrication 2015, 7 (4), 045009. DOI: 10.1088/1758-5090/7/4/045009.
26
27
28 [77] Lin, H.; Tang, Y.; Lozito, T. P.; Oyster, N.; Kang, R. B.; Fritch, M. R.; Wang, B.; Tuan, R. S.,
29
30
Projection Stereolithographic Fabrication of BMP-2 Gene-activated Matrix for Bone Tissue
31
32
33 Engineering. Scientific reports 2017, 7 (1), 11327. DOI: 10.1038/s41598-017-11051-0.
34
35 [78] Zhou, X.; Castro, N. J.; Zhu, W.; Cui, H.; Aliabouzar, M.; Sarkar, K.; Zhang, L. G., Improved
36
37
38 Human Bone Marrow Mesenchymal Stem Cell Osteogenesis in 3D Bioprinted Tissue Scaffolds with
39
40 Low Intensity Pulsed Ultrasound Stimulation. Scientific reports 2016, 6, 32876. DOI:
41
42
43 10.1038/srep32876.
44
45 [79] Castro, N. J.; O'Brien, J.; Zhang, L. G., Integrating biologically inspired nanomaterials and
46
47
table-top stereolithography for 3D printed biomimetic osteochondral scaffolds. Nanoscale 2015, 7 (33),
48
49
50 14010-22. DOI: 10.1039/c5nr03425f.
51
52
[80] Zhu, W.; Holmes, B.; Glazer, R. I.; Zhang, L. G., 3D printed nanocomposite matrix for the study
53
54
55 of breast cancer bone metastasis. Nanomedicine : nanotechnology, biology, and medicine 2016, 12 (1),
56
57
53
58
59
1
2
3
69-79. DOI: 10.1016/j.nano.2015.09.010.
4
5
6 [81] Blanquer, S. B. G.; Gebraad, A. W. H.; Miettinen, S.; Poot, A. A.; Grijpma, D. W.; Haimi, S. P.,
7
8
Differentiation of adipose stem cells seeded towards annulus fibrosus cells on a designed
9
10
11 poly(trimethylene carbonate) scaffold prepared by stereolithography. Journal of tissue engineering and
12
13 regenerative medicine 2017, 11 (10), 2752-2762. DOI: 10.1002/term.2170.
14
15
16 [82] Wei, Z.; Harris, B. T.; Zhang, L. G., Gelatin methacrylamide hydrogel with graphene nanoplatelets
17
18 for neural cell-laden 3D bioprinting. Conference proceedings : ... Annual International Conference of
19
20
21 the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology
22
23 Society. Annual Conference 2016, 2016, 4185-4188. DOI: 10.1109/EMBC.2016.7591649.
24
25
[83] Bajaj, P.; Marchwiany, D.; Duarte, C.; Bashir, R., Patterned three-dimensional encapsulation of
26
27
28 embryonic stem cells using dielectrophoresis and stereolithography. Advanced healthcare materials
29
30
2013, 2 (3), 450-8. DOI: 10.1002/adhm.201200318.
31
32
33 [84] Arcaute, K.; Mann, B.; Wicker, R., Stereolithography of spatially controlled multi-material
34
35 bioactive poly(ethylene glycol) scaffolds. Acta biomaterialia 2010, 6 (3), 1047-54. DOI:
36
37
38 10.1016/j.actbio.2009.08.017.
39
40 [85] Chan, V.; Zorlutuna, P.; Jeong, J. H.; Kong, H.; Bashir, R., Three-dimensional photopatterning of
41
42
43 hydrogels using stereolithography for long-term cell encapsulation. Lab on a chip 2010, 10 (16),
44
45 2062-70. DOI: 10.1039/c004285d.
46
47
[86] Hong, S.; Lee, J. Y.; Hwang, C.; Shin, J. H.; Park, Y., Inhibition of Rho-Associated Protein Kinase
48
49
50 Increases the Angiogenic Potential of Mesenchymal Stem Cell Aggregates via Paracrine Effects. Tissue
51
52
engineering. Part A 2016, 22 (3-4), 233-43. DOI: 10.1089/ten.TEA.2015.0289.
53
54
55 [87] Coburn, L.; Lopez, H.; Caldwell, B. J.; Moussa, E.; Yap, C.; Priya, R.; Noppe, A.; Roberts, A. P.;
56
57
54
58
59
1
2
3
Lobaskin, V.; Yap, A. S.; Neufeld, Z.; Gomez, G. A., Contact inhibition of locomotion and mechanical
4
5
6 cross-talk between cell-cell and cell-substrate adhesion determine the pattern of junctional tension in
7
8
epithelial cell aggregates. Molecular biology of the cell 2016, 27 (22), 3436-3448. DOI:
9
10
11 10.1091/mbc.E16-04-0226.
12
13 [88] Zimmermann, J. A.; McDevitt, T. C., Pre-conditioning mesenchymal stromal cell spheroids for
14
15
16 immunomodulatory paracrine factor secretion. Cytotherapy 2014, 16 (3), 331-45. DOI:
17
18 10.1016/j.jcyt.2013.09.004.
19
20
21 [89] Komatsu, M.; Konagaya, S.; Egawa, E. Y.; Iwata, H., Maturation of human iPS cell-derived
22
23 dopamine neuron precursors in alginate-Ca(2+) hydrogel. Biochimica et biophysica acta 2015, 1850
24
25
(9), 1669-75. DOI: 10.1016/j.bbagen.2015.04.011.
26
27
28 [90] Lei, J.; Trevino, E.; Temenoff, J., Cell number and chondrogenesis in human mesenchymal stem
29
30
cell aggregates is affected by the sulfation level of heparin used as a cell coating. Journal of biomedical
31
32
33 materials research. Part A 2016, 104 (7), 1817-29. DOI: 10.1002/jbm.a.35713.
34
35 [91] Lin, R. Z.; Chang, H. Y., Recent advances in three-dimensional multicellular spheroid culture for
36
37
38 biomedical research. Biotechnology journal 2008, 3 (9-10), 1172-84. DOI: 10.1002/biot.200700228.
39
40 [92] Marga, F.; Jakab, K.; Khatiwala, C.; Shepherd, B.; Dorfman, S.; Hubbard, B.; Colbert, S.; Gabor,
41
42
43 F., Toward engineering functional organ modules by additive manufacturing. Biofabrication 2012, 4 (2),
44
45 022001. DOI: 10.1088/1758-5082/4/2/022001.
46
47
[93] Marchioli, G.; van Gurp, L.; van Krieken, P. P.; Stamatialis, D.; Engelse, M.; van Blitterswijk, C.
48
49
50 A.; Karperien, M. B.; de Koning, E.; Alblas, J.; Moroni, L.; van Apeldoorn, A. A., Fabrication of
51
52
three-dimensional bioplotted hydrogel scaffolds for islets of Langerhans transplantation. Biofabrication
53
54
55 2015, 7 (2), 025009. DOI: 10.1088/1758-5090/7/2/025009.
56
57
55
58
59
1
2
3
[94] Tan, Y.; Richards, D. J.; Trusk, T. C.; Visconti, R. P.; Yost, M. J.; Kindy, M. S.; Drake, C. J.;
4
5
6 Argraves, W. S.; Markwald, R. R.; Mei, Y., 3D printing facilitated scaffold-free tissue unit fabrication.
7
8
Biofabrication 2014, 6 (2), 024111. DOI: 10.1088/1758-5082/6/2/024111.
9
10
11 [95] Yang, X.; Sun, Y.; Wang, Q., A phase field approach for multicellular aggregate fusion in
12
13 biofabrication. Journal of biomechanical engineering 2013, 135 (7), 71005. DOI: 10.1115/1.4024139.
14
15
16 [96] Yang, X.; Mironov, V.; Wang, Q., Modeling fusion of cellular aggregates in biofabrication using
17
18 phase field theories. Journal of theoretical biology 2012, 303, 110-8. DOI: 10.1016/j.jtbi.2012.03.003.
19
20
21 [97] Mironov, V.; Visconti, R. P.; Kasyanov, V.; Forgacs, G.; Drake, C. J.; Markwald, R. R., Organ
22
23 printing: tissue spheroids as building blocks. Biomaterials 2009, 30 (12), 2164-74. DOI:
24
25
10.1016/j.biomaterials.2008.12.084.
26
27
28 [98] Riba, J.; Renz, N.; Niemoller, C.; Bleul, S.; Pfeifer, D.; Stosch, J. M.; Metzeler, K. H.; Hackanson,
29
30
B.; Lubbert, M.; Duyster, J.; Koltay, P.; Zengerle, R.; Claus, R.; Zimmermann, S.; Becker, H.,
31
32
33 Molecular Genetic Characterization of Individual Cancer Cells Isolated via Single-Cell Printing. PloS
34
35 one 2016, 11 (9), e0163455. DOI: 10.1371/journal.pone.0163455.
36
37
38 [99] Stumpf, F.; Schoendube, J.; Gross, A.; Rath, C.; Niekrawietz, S.; Koltay, P.; Roth, G., Single-cell
39
40 PCR of genomic DNA enabled by automated single-cell printing for cell isolation. Biosensors &
41
42
43 bioelectronics 2015, 69, 301-6. DOI: 10.1016/j.bios.2015.03.008.
44
45 [100] Barron, J. A.; Krizman, D. B.; Ringeisen, B. R., Laser printing of single cells: statistical analysis,
46
47
cell viability, and stress. Annals of biomedical engineering 2005, 33 (2), 121-30.
48
49
50 [101] Zhang, K.; Chou, C. K.; Xia, X.; Hung, M. C.; Qin, L., Block-Cell-Printing for live single-cell
51
52
printing. Proceedings of the National Academy of Sciences of the United States of America 2014, 111
53
54
55 (8), 2948-53. DOI: 10.1073/pnas.1313661111.
56
57
56
58
59
1
2
3
[102] Benning, L.; Gutzweiler, L.; Trondle, K.; Riba, J.; Zengerle, R.; Koltay, P.; Zimmermann, S.;
4
5
6 Stark, G. B.; Finkenzeller, G., Cytocompatibility testing of hydrogels toward bioprinting of
7
8
mesenchymal stem cells. Journal of biomedical materials research. Part A 2017, 105 (12), 3231-3241.
9
10
11 DOI: 10.1002/jbm.a.36179.
12
13 [103] Guvendiren M.; Lu H. D.; Burdick J. A., Shear-thinning hydrogels for biomedical applications[J].
14
15
16 Soft matter, 2012, 8(2): 260-272.
17
18 [104] Skardal, A.; Mack, D.; Kapetanovic, E.; Atala, A.; Jackson, J. D.; Yoo, J.; Soker, S., Bioprinted
19
20
21 amniotic fluid-derived stem cells accelerate healing of large skin wounds. Stem cells translational
22
23 medicine 2012, 1 (11), 792-802. DOI: 10.5966/sctm.2012-0088.
24
25
[105] Lin, L.; Chu, Y. S.; Thiery, J. P.; Lim, C. T.; Rodriguez, I., Microfluidic cell trap array for
26
27
28 controlled positioning of single cells on adhesive micropatterns. Lab on a chip 2013, 13 (4), 714-21.
29
30
DOI: 10.1039/c2lc41070b.
31
32
33 [106] Zychowicz, M.; Mehn, D.; Ruiz, A.; Frontczak-Baniewicz, M.; Rossi, F.; Buzanska, L.,
34
35 Patterning of human cord blood-derived stem cells on single cell posts and lines: Implications for
36
37
38 neural commitment. Acta neurobiologiae experimentalis 2012, 72 (4), 325-36.
39
40 [107] Mousavi, S. J.; Doweidar, M. H., Role of Mechanical Cues in Cell Differentiation and
41
42
43 Proliferation: A 3D Numerical Model. PloS one 2015, 10 (5), e0124529. DOI:
44
45 10.1371/journal.pone.0124529.
46
47
[108] Lessey, E. C.; Guilluy, C.; Burridge, K., From mechanical force to RhoA activation. Biochemistry
48
49
50 2012, 51 (38), 7420-32. DOI: 10.1021/bi300758e.
51
52
[109] Blaeser, A.; Duarte Campos, D. F.; Puster, U.; Richtering, W.; Stevens, M. M.; Fischer, H.,
53
54
55 Controlling Shear Stress in 3D Bioprinting is a Key Factor to Balance Printing Resolution and Stem
56
57
57
58
59
1
2
3
Cell Integrity. Advanced healthcare materials 2016, 5 (3), 326-33. DOI: 10.1002/adhm.201500677.
4
5
6 [110] Aguado, B. A.; Mulyasasmita, W.; Su, J.; Lampe, K. J.; Heilshorn, S. C., Improving viability of
7
8
stem cells during syringe needle flow through the design of hydrogel cell carriers. Tissue engineering.
9
10
11 Part A 2012, 18 (7-8), 806-15. DOI: 10.1089/ten.TEA.2011.0391.
12
13 [111] Skardal, A.; Atala, A., Biomaterials for integration with 3-D bioprinting. Annals of biomedical
14
15
16 engineering 2015, 43 (3), 730-46. DOI: 10.1007/s10439-014-1207-1.
17
18 [112] Nair, K.; Gandhi, M.; Khalil, S.; Yan, K. C.; Marcolongo, M.; Barbee, K.; Sun, W.,
19
20
21 Characterization of cell viability during bioprinting processes. Biotechnology journal 2009, 4 (8),
22
23 1168-77. DOI: 10.1002/biot.200900004.
24
25
[113] Kang, K. H.; Hockaday, L. A.; Butcher, J. T., Quantitative optimization of solid freeform
26
27
28 deposition of aqueous hydrogels. Biofabrication 2013, 5 (3), 035001. DOI:
29
30
10.1088/1758-5082/5/3/035001.
31
32
33 [114] Raof, N. A.; Schiele, N. R.; Xie, Y.; Chrisey, D. B.; Corr, D. T., The maintenance of pluripotency
34
35 following laser direct-write of mouse embryonic stem cells. Biomaterials 2011, 32 (7), 1802-8. DOI:
36
37
38 10.1016/j.biomaterials.2010.11.015.
39
40 [115] Markstedt, K.; Mantas, A.; Tournier, I.; Martinez Avila, H.; Hagg, D.; Gatenholm, P., 3D
41
42
43 Bioprinting Human Chondrocytes with Nanocellulose-Alginate Bioink for Cartilage Tissue
44
45 Engineering Applications. Biomacromolecules 2015, 16 (5), 1489-96. DOI:
46
47
10.1021/acs.biomac.5b00188.
48
49
50 [116] Thakur, A.; Jaiswal, M. K.; Peak, C. W.; Carrow, J. K.; Gentry, J.; Dolatshahi-Pirouz, A.;
51
52
Gaharwar, A. K., Injectable shear-thinning nanoengineered hydrogels for stem cell delivery. Nanoscale
53
54
55 2016, 8 (24), 12362-72. DOI: 10.1039/c6nr02299e.
56
57
58
58
59
1
2
3
[117] Ouyang, L.; Highley, C. B.; Sun, W.; Burdick, J. A., A Generalizable Strategy for the 3D
4
5
6 Bioprinting of Hydrogels from Nonviscous Photo-crosslinkable Inks. Advanced materials 2017, 29 (8).
7
8
DOI: 10.1002/adma.201604983.
9
10
11 [118] Colosi, C.; Shin, S. R.; Manoharan, V.; Massa, S.; Costantini, M.; Barbetta, A.; Dokmeci, M. R.;
12
13 Dentini, M.; Khademhosseini, A., Microfluidic Bioprinting of Heterogeneous 3D Tissue Constructs
14
15
16 Using Low-Viscosity Bioink. Advanced materials 2016, 28 (4), 677-84. DOI:
17
18 10.1002/adma.201503310.
19
20
21 [119] Snyder, J.; Rin Son, A.; Hamid, Q.; Wang, C.; Lui, Y.; Sun, W., Mesenchymal stem cell printing
22
23 and process regulated cell properties. Biofabrication 2015, 7 (4), 044106. DOI:
24
25
10.1088/1758-5090/7/4/044106.
26
27
28 [120] Nahmias, Y.; Odde, D. J., Micropatterning of living cells by laser-guided direct writing:
29
30
application to fabrication of hepatic-endothelial sinusoid-like structures. Nature protocols 2006, 1 (5),
31
32
33 2288-96. DOI: 10.1038/nprot.2006.386.
34
35 [121] Xu, T.; Gregory, C. A.; Molnar, P.; Cui, X.; Jalota, S.; Bhaduri, S. B.; Boland, T., Viability and
36
37
38 electrophysiology of neural cell structures generated by the inkjet printing method. Biomaterials 2006,
39
40 27 (19), 3580-8. DOI: 10.1016/j.biomaterials.2006.01.048.
41
42
43 [122] Schiele, N. R.; Corr, D. T.; Huang, Y.; Raof, N. A.; Xie, Y.; Chrisey, D. B., Laser-based
44
45 direct-write techniques for cell printing. Biofabrication 2010, 2 (3), 032001. DOI:
46
47
10.1088/1758-5082/2/3/032001.
48
49
50 [123] Wust, S.; Godla, M. E.; Muller, R.; Hofmann, S., Tunable hydrogel composite with two-step
51
52
processing in combination with innovative hardware upgrade for cell-based three-dimensional
53
54
55 bioprinting. Acta biomaterialia 2014, 10 (2), 630-40. DOI: 10.1016/j.actbio.2013.10.016.
56
57
59
58
59
1
2
3
[124] Ren, X.; Ott, H. C., On the road to bioartificial organs. Pflugers Archiv : European journal of
4
5
6 physiology 2014, 466 (10), 1847-57. DOI: 10.1007/s00424-014-1504-4.
7
8
[125] Wang, X. F.; Song, Y.; Liu, Y. S.; Sun, Y. C.; Wang, Y. G.; Wang, Y.; Lyu, P. J., Osteogenic
9
10
11 Differentiation of Three-Dimensional Bioprinted Constructs Consisting of Human Adipose-Derived
12
13 Stem Cells In Vitro and In Vivo. PloS one 2016, 11 (6), e0157214. DOI:
14
15
16 10.1371/journal.pone.0157214.
17
18 [126] Fedorovich, N. E.; Alblas, J.; de Wijn, J. R.; Hennink, W. E.; Verbout, A. J.; Dhert, W. J.,
19
20
21 Hydrogels as extracellular matrices for skeletal tissue engineering: state-of-the-art and novel
22
23 application in organ printing. Tissue engineering 2007, 13 (8), 1905-25. DOI: 10.1089/ten.2006.0175.
24
25
[127] Rutz, A. L.; Hyland, K. E.; Jakus, A. E.; Burghardt, W. R.; Shah, R. N., A multimaterial bioink
26
27
28 method for 3D printing tunable, cell-compatible hydrogels. Advanced materials 2015, 27 (9), 1607-14.
29
30
DOI: 10.1002/adma.201405076.
31
32
33 [128] Wells, R. G., The role of matrix stiffness in regulating cell behavior. Hepatology 2008, 47 (4),
34
35 1394-400. DOI: 10.1002/hep.22193.
36
37
38 [129] Hwang, J. H.; Byun, M. R.; Kim, A. R.; Kim, K. M.; Cho, H. J.; Lee, Y. H.; Kim, J.; Jeong, M. G.;
39
40 Hwang, E. S.; Hong, J. H., Extracellular Matrix Stiffness Regulates Osteogenic Differentiation through
41
42
43 MAPK Activation. PloS one 2015, 10 (8), e0135519. DOI: 10.1371/journal.pone.0135519.
44
45 [130] Huang, Q.; Elkhooly, T. A.; Liu, X.; Zhang, R.; Yang, X.; Shen, Z.; Feng, Q., Effects of
46
47
hierarchical micro/nano-topographies on the morphology, proliferation and differentiation of
48
49
50 osteoblast-like cells. Colloids and surfaces. B, Biointerfaces 2016, 145, 37-45. DOI:
51
52
10.1016/j.colsurfb.2016.04.031.
53
54
55 [131] Dolatshahi-Pirouz, A.; Pennisi, C. P.; Skeldal, S.; Foss, M.; Chevallier, J.; Zachar, V.; Andreasen,
56
57
60
58
59
1
2
3
P.; Yoshida, K.; Besenbacher, F., The influence of glancing angle deposited nano-rough platinum
4
5
6 surfaces on the adsorption of fibrinogen and the proliferation of primary human fibroblasts.
7
8
Nanotechnology 2009, 20 (9), 095101. DOI: 10.1088/0957-4484/20/9/095101.
9
10
11 [132] Lin, X.; Shi, Y.; Cao, Y.; Liu, W., Recent progress in stem cell differentiation directed by material
12
13 and mechanical cues. Biomedical materials 2016, 11 (1), 014109. DOI:
14
15
16 10.1088/1748-6041/11/1/014109.
17
18 [133]Poellmann, M. J.; Johnson, A. J., Multimaterial polyacrylamide: fabrication with
19
20
21 electrohydrodynamic jet printing, applications, and modeling. Biofabrication 2014, 6 (3), 035018. DOI:
22
23 10.1088/1758-5082/6/3/035018.
24
25
[134] Kobelt, L. J.; Wilkinson, A. E.; McCormick, A. M.; Willits, R. K.; Leipzig, N. D., Short duration
26
27
28 electrical stimulation to enhance neurite outgrowth and maturation of adult neural stem progenitor cells.
29
30
Annals of biomedical engineering 2014, 42 (10), 2164-76. DOI: 10.1007/s10439-014-1058-9.
31
32
33 [135] Duarte Campos, D. F.; Blaeser, A.; Korsten, A.; Neuss, S.; Jakel, J.; Vogt, M.; Fischer, H., The
34
35 stiffness and structure of three-dimensional printed hydrogels direct the differentiation of mesenchymal
36
37
38 stromal cells toward adipogenic and osteogenic lineages. Tissue engineering. Part A 2015, 21 (3-4),
39
40 740-56. DOI: 10.1089/ten.TEA.2014.0231.
41
42
43 [136] Barnes, C. A.; Brison, J.; Michel, R.; Brown, B. N.; Castner, D. G.; Badylak, S. F.; Ratner, B. D.,
44
45 The surface molecular functionality of decellularized extracellular matrices. Biomaterials 2011, 32 (1),
46
47
137-43. DOI: 10.1016/j.biomaterials.2010.09.007.
48
49
50 [137] Pati, F.; Jang, J.; Ha, D. H.; Won Kim, S.; Rhie, J. W.; Shim, J. H.; Kim, D. H.; Cho, D. W.,
51
52
Printing three-dimensional tissue analogues with decellularized extracellular matrix bioink. Nature
53
54
55 communications 2014, 5, 3935. DOI: 10.1038/ncomms4935.
56
57
61
58
59
1
2
3
[138] Pati, F.; Ha, D. H.; Jang, J.; Han, H. H.; Rhie, J. W.; Cho, D. W., Biomimetic 3D tissue printing
4
5
6 for soft tissue regeneration. Biomaterials 2015, 62, 164-75. DOI: 10.1016/j.biomaterials.2015.05.043.
7
8
[139] Zhang, Y.; He, Y.; Bharadwaj, S.; Hammam, N.; Carnagey, K.; Myers, R.; Atala, A.; Van Dyke,
9
10
11 M., Tissue-specific extracellular matrix coatings for the promotion of cell proliferation and
12
13 maintenance of cell phenotype. Biomaterials 2009, 30 (23-24), 4021-8. DOI:
14
15
16 10.1016/j.biomaterials.2009.04.005.
17
18 [140] Li, C.; Faulkner-Jones, A.; Dun, A. R.; Jin, J.; Chen, P.; Xing, Y.; Yang, Z.; Li, Z.; Shu, W.; Liu,
19
20
21 D.; Duncan, R. R., Rapid formation of a supramolecular polypeptide-DNA hydrogel for in situ
22
23 three-dimensional multilayer bioprinting. Angewandte Chemie 2015, 54 (13), 3957-61. DOI:
24
25
10.1002/anie.201411383.
26
27
28 [141] Marcu, I. C.; Illaste, A.; Heuking, P.; Jaconi, M. E.; Ullrich, N. D., Functional Characterization
29
30
and Comparison of Intercellular Communication in Stem Cell-Derived Cardiomyocytes. Stem cells
31
32
33 2015, 33 (7), 2208-18. DOI: 10.1002/stem.2009.
34
35 [142] Tang, J.; Peng, R.; Ding, J., The regulation of stem cell differentiation by cell-cell contact on
36
37
38 micropatterned material surfaces. Biomaterials 2010, 31 (9), 2470-6. DOI:
39
40 10.1016/j.biomaterials.2009.12.006.
41
42
43 [143] Xu, F.; Sridharan, B.; Wang, S.; Gurkan, U. A.; Syverud, B.; Demirci, U., Embryonic stem cell
44
45 bioprinting for uniform and controlled size embryoid body formation. Biomicrofluidics 2011, 5 (2),
46
47
22207. DOI: 10.1063/1.3580752.
48
49
50 [144] Ouyang, L.; Yao, R.; Mao, S.; Chen, X.; Na, J.; Sun, W., Three-dimensional bioprinting of
51
52
embryonic stem cells directs highly uniform embryoid body formation. Biofabrication 2015, 7 (4),
53
54
55 044101. DOI: 10.1088/1758-5090/7/4/044101.
56
57
62
58
59
1
2
3
[145] Wang, Z.; Xia, J.; Yan, Y.; Tsai, A. C.; Li, Y.; Ma, T.; Guan, J., Facile functionalization and
4
5
6 assembly of live cells with microcontact-printed polymeric biomaterials. Acta biomaterialia 2015, 11,
7
8
80-7. DOI: 10.1016/j.actbio.2014.10.006.
9
10
11 [146] Roy, N. S.; Cleren, C.; Singh, S. K.; Yang, L.; Beal, M. F.; Goldman, S. A., Functional
12
13 engraftment of human ES cell-derived dopaminergic neurons enriched by coculture with
14
15
16 telomerase-immortalized midbrain astrocytes. Nature medicine 2006, 12 (11), 1259-68. DOI:
17
18 10.1038/nm1495.
19
20
21 [147] Yao, R.; Zhang, R.; Lin, F.; Luan, J., Biomimetic injectable
22
23 HUVEC-adipocytes/collagen/alginate microsphere co-cultures for adipose tissue engineering.
24
25
Biotechnology and bioengineering 2013, 110 (5), 1430-43. DOI: 10.1002/bit.24784.
26
27
28 [148] Bourget, J. M.; Kerouredan, O.; Medina, M.; Remy, M.; Thebaud, N. B.; Bareille, R.; Chassande,
29
30
O.; Amedee, J.; Catros, S.; Devillard, R., Patterning of Endothelial Cells and Mesenchymal Stem Cells
31
32
33 by Laser-Assisted Bioprinting to Study Cell Migration. BioMed research international 2016, 2016,
34
35 3569843. DOI: 10.1155/2016/3569843.
36
37
38 [149] Kolesky, D. B.; Truby, R. L.; Gladman, A. S.; Busbee, T. A.; Homan, K. A.; Lewis, J. A., 3D
39
40 bioprinting of vascularized, heterogeneous cell-laden tissue constructs. Advanced materials 2014, 26
41
42
43 (19), 3124-30. DOI: 10.1002/adma.201305506.
44
45 [150] Wang, X.; Schroder, H. C.; Muller, W. E., Enzymatically synthesized inorganic polymers as
46
47
morphogenetically active bone scaffolds: application in regenerative medicine. International review of
48
49
50 cell and molecular biology 2014, 313, 27-77. DOI: 10.1016/B978-0-12-800177-6.00002-5.
51
52
[151] Engler, A. J.; Sen, S.; Sweeney, H. L.; Discher, D. E., Matrix elasticity directs stem cell lineage
53
54
55 specification. Cell 2006, 126 (4), 677-89. DOI: 10.1016/j.cell.2006.06.044.
56
57
63
58
59
1
2
3
[152] Tong, X.; Yang, F., Sliding Hydrogels with Mobile Molecular Ligands and Crosslinks as 3D
4
5
6 Stem Cell Niche. Advanced materials 2016, 28 (33), 7257-63. DOI: 10.1002/adma.201601484.
7
8
[153] Poldervaart, M. T.; Wang, H.; van der Stok, J.; Weinans, H.; Leeuwenburgh, S. C.; Oner, F. C.;
9
10
11 Dhert, W. J.; Alblas, J., Sustained release of BMP-2 in bioprinted alginate for osteogenicity in mice and
12
13 rats. PloS one 2013, 8 (8), e72610. DOI: 10.1371/journal.pone.0072610.
14
15
16 [154] Ker, E. D.; Chu, B.; Phillippi, J. A.; Gharaibeh, B.; Huard, J.; Weiss, L. E.; Campbell, P. G.,
17
18 Engineering spatial control of multiple differentiation fates within a stem cell population. Biomaterials
19
20
21 2011, 32 (13), 3413-22. DOI: 10.1016/j.biomaterials.2011.01.036.
22
23 [155] Lutolf, M. P.; Hubbell, J. A., Synthetic biomaterials as instructive extracellular
24
25
microenvironments for morphogenesis in tissue engineering. Nature biotechnology 2005, 23 (1), 47-55.
26
27
28 DOI: 10.1038/nbt1055.
29
30
[156] West J. L.; Hubbell J. A., Polymeric biomaterials with degradation sites for proteases involved in
31
32
33 cell migration. Macromolecules, 1999, 32(1): 241-244.
34
35 [157] Fedorovich, N. E.; Kuipers, E.; Gawlitta, D.; Dhert, W. J.; Alblas, J., Scaffold porosity and
36
37
38 oxygenation of printed hydrogel constructs affect functionality of embedded osteogenic progenitors.
39
40 Tissue engineering. Part A 2011, 17 (19-20), 2473-86. DOI: 10.1089/ten.TEA.2011.0001.
41
42
43 [158] Arslan-Yildiz, A.; El Assal, R.; Chen, P.; Guven, S.; Inci, F.; Demirci, U., Towards artificial tissue
44
45 models: past, present, and future of 3D bioprinting. Biofabrication 2016, 8 (1), 014103. DOI:
46
47
10.1088/1758-5090/8/1/014103.
48
49
50 [159] Domingos, M.; Intranuovo, F.; Russo, T.; De Santis, R.; Gloria, A.; Ambrosio, L.; Ciurana, J.;
51
52
Bartolo, P., The first systematic analysis of 3D rapid prototyped poly(epsilon-caprolactone) scaffolds
53
54
55 manufactured through BioCell printing: the effect of pore size and geometry on compressive
56
57
64
58
59
1
2
3
mechanical behaviour and in vitro hMSC viability. Biofabrication 2013, 5 (4), 045004. DOI:
4
5
6 10.1088/1758-5082/5/4/045004.
7
8
[160] Hillig, W. B.; Choi, Y.; Murthy, S.; Natravali, N.; Ajayan, P., An open-pored
9
10
11 gelatin/hydroxyapatite composite as a potential bone substitute. Journal of materials science. Materials
12
13 in medicine 2008, 19 (1), 11-7. DOI: 10.1007/s10856-007-0154-x.
14
15
16 [161] Rouwkema, J.; Rivron, N. C.; van Blitterswijk, C. A., Vascularization in tissue engineering.
17
18 Trends in biotechnology 2008, 26 (8), 434-41. DOI: 10.1016/j.tibtech.2008.04.009.
19
20
21 [162] Kaully, T.; Kaufman-Francis, K.; Lesman, A.; Levenberg, S., Vascularization--the conduit to
22
23 viable engineered tissues. Tissue engineering. Part B, Reviews 2009, 15 (2), 159-69. DOI:
24
25
10.1089/ten.teb.2008.0193.
26
27
28 [163] Richards, D.; Jia, J.; Yost, M.; Markwald, R.; Mei, Y., 3D Bioprinting for Vascularized Tissue
29
30
Fabrication. Annals of biomedical engineering 2017, 45 (1), 132-147. DOI:
31
32
33 10.1007/s10439-016-1653-z.
34
35 [164] Udan, R. S.; Culver, J. C.; Dickinson, M. E., Understanding vascular development. Wiley
36
37
38 interdisciplinary reviews. Developmental biology 2013, 2 (3), 327-46. DOI: 10.1002/wdev.91.
39
40 [165] Kolesky, D. B.; Homan, K. A.; Skylar-Scott, M. A.; Lewis, J. A., Three-dimensional bioprinting
41
42
43 of thick vascularized tissues. Proceedings of the National Academy of Sciences of the United States of
44
45 America 2016, 113 (12), 3179-84. DOI: 10.1073/pnas.1521342113.
46
47
[166] Datta, P.; Ayan, B.; Ozbolat, I. T., Bioprinting for vascular and vascularized tissue biofabrication.
48
49
50 Acta biomaterialia 2017, 51, 1-20. DOI: 10.1016/j.actbio.2017.01.035.
51
52
[167] Chung, J. C.; Shum-Tim, D., Neovascularization in tissue engineering. Cells 2012, 1 (4), 1246-60.
53
54
55 DOI: 10.3390/cells1041246.
56
57
65
58
59
1
2
3
[168] Baldwin, J.; Antille, M.; Bonda, U.; De-Juan-Pardo, E. M.; Khosrotehrani, K.; Ivanovski, S.;
4
5
6 Petcu, E. B.; Hutmacher, D. W., In vitro pre-vascularisation of tissue-engineered constructs A
7
8
co-culture perspective. Vascular cell 2014, 6, 13. DOI: 10.1186/2045-824X-6-13.
9
10
11 [169] Jang, J.; Park, J. Y.; Gao, G.; Cho, D. W., Biomaterials-based 3D cell printing for next-generation
12
13 therapeutics and diagnostics. Biomaterials 2018, 156, 88-106. DOI:
14
15
16 10.1016/j.biomaterials.2017.11.030.
17
18 [170] Gopinathan, J.; Noh, I., Recent trends in bioinks for 3D printing. Biomaterials research 2018, 22,
19
20
21 11. DOI: 10.1186/s40824-018-0122-1.
22
23 [171] Khetan, S.; Guvendiren, M.; Legant, W. R.; Cohen, D. M.; Chen, C. S.; Burdick, J. A.,
24
25
Degradation-mediated cellular traction directs stem cell fate in covalently crosslinked
26
27
28 three-dimensional hydrogels. Nature materials 2013, 12 (5), 458-65. DOI: 10.1038/nmat3586.
29
30
[172] Ma, Y.; Thiele, J.; Abdelmohsen, L.; Xu, J.; Huck, W. T., Biocompatible macro-initiators
31
32
33 controlling radical retention in microfluidic on-chip photo-polymerization of water-in-oil emulsions.
34
35 Chemical communications 2014, 50 (1), 112-4. DOI: 10.1039/c3cc46733c.
36
37
38 [173] Zhang, C.; Wen, X.; Vyavahare, N. R.; Boland, T., Synthesis and characterization of
39
40 biodegradable elastomeric polyurethane scaffolds fabricated by the inkjet technique. Biomaterials 2008,
41
42
43 29 (28), 3781-91. DOI: 10.1016/j.biomaterials.2008.06.009.
44
45 [174] Lee, H. J.; Kim, Y. B.; Ahn, S. H.; Lee, J. S.; Jang, C. H.; Yoon, H.; Chun, W.; Kim, G. H., A
46
47
New Approach for Fabricating Collagen/ECM-Based Bioinks Using Preosteoblasts and Human
48
49
50 Adipose Stem Cells. Advanced healthcare materials 2015, 4 (9), 1359-68. DOI:
51
52
10.1002/adhm.201500193.
53
54
55 [175] Ye, K.; Felimban, R.; Traianedes, K.; Moulton, S. E.; Wallace, G. G.; Chung, J.; Quigley, A.;
56
57
66
58
59
1
2
3
Choong, P. F.; Myers, D. E., Chondrogenesis of infrapatellar fat pad derived adipose stem cells in 3D
4
5
6 printed chitosan scaffold. PloS one 2014, 9 (6), e99410. DOI: 10.1371/journal.pone.0099410.
7
8
[176] Hong, S.; Song, S. J.; Lee, J. Y.; Jang, H.; Choi, J.; Sun, K.; Park, Y., Cellular behavior in
9
10
11 micropatterned hydrogels by bioprinting system depended on the cell types and cellular interaction.
12
13 Journal of bioscience and bioengineering 2013, 116 (2), 224-30. DOI: 10.1016/j.jbiosc.2013.02.011.
14
15
16 [177] Gao, G.; Yonezawa, T.; Hubbell, K.; Dai, G.; Cui, X., Inkjet-bioprinted acrylated peptides and
17
18 PEG hydrogel with human mesenchymal stem cells promote robust bone and cartilage formation with
19
20
21 minimal printhead clogging. Biotechnology journal 2015, 10 (10), 1568-77. DOI:
22
23 10.1002/biot.201400635.
24
25
[178] Merceron, T. K.; Burt, M.; Seol, Y. J.; Kang, H. W.; Lee, S. J.; Yoo, J. J.; Atala, A., A 3D
26
27
28 bioprinted complex structure for engineering the muscle-tendon unit. Biofabrication 2015, 7 (3),
29
30
035003. DOI: 10.1088/1758-5090/7/3/035003.
31
32
33 [179] Sun, Y.; Yang, X.; Wang, Q., In-silico analysis on biofabricating vascular networks using kinetic
34
35 Monte Carlo simulations. Biofabrication 2014, 6 (1), 015008. DOI: 10.1088/1758-5082/6/1/015008.
36
37
38 [180] McCune, M.; Shafiee, A.; Forgacs, G.; Kosztin, I., Predictive modeling of post bioprinting
39
40 structure formation. Soft matter 2014, 10 (11), 1790-800.
41
42
43 [181] Mironov, V.; Kasyanov, V.; Markwald, R. R., Organ printing: from bioprinter to organ
44
45 biofabrication line. Current opinion in biotechnology 2011, 22 (5), 667-73. DOI:
46
47
10.1016/j.copbio.2011.02.006.
48
49
50 [182] Rimann, M.; Bono, E.; Annaheim, H.; Bleisch, M.; Graf-Hausner, U., Standardized 3D
51
52
Bioprinting of Soft Tissue Models with Human Primary Cells. Journal of laboratory automation 2016,
53
54
55 21 (4), 496-509. DOI: 10.1177/2211068214567146.
56
57
67
58
59
1
2
3
[183] Ojeh, N.; Pastar, I.; Tomic-Canic, M.; Stojadinovic, O., Stem Cells in Skin Regeneration, Wound
4
5
6 Healing, and Their Clinical Applications. International journal of molecular sciences 2015, 16 (10),
7
8
25476-501. DOI: 10.3390/ijms161025476.
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
68
58
59
1
2
3
4 For Table of Contents Use Only
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
Manuscript title: "Bioprinting of stem cells: interplay
21
22 of bioprinting process, bioinks and stem cell proper
23
24
ties."
25
26
27
28
29 Author(s): Ding, Supeng; Feng, Lu; Wu, Jiayang; Zhu,
30
31
32 Fei; Tan, Ze'en; Yao, Rui
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
69
58
59
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30 Figure 1. Schematic of the whole lifespan of stem cell bioprinting practice. Reproduced with permission from
ref 27,28. Copyright Biofabrication.
31
32 1763x1234mm (72 x 72 DPI)
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45 Figure 2. Illustrations and examples of different bioprinting techniques. (A) Multi-cell bioprinting, which can
46 be further divided into inkjet bioprinting (image from MicroFab Technologies, Inc.), microextrusion-based
47 bioprinting (image from Organovo Holdings, Inc.), laser-assisted bioprinting (image from Organovo
48 Holdings, Inc.) and stereolithography-based bioprinting (image from Formlabs, Inc.). (B) Cell aggregate
49 bioprinting. Reproduced with permission from ref 30. Copyright Biomaterials. (C) Single-cell bioprinting.
Reproduced with permission from ref 31. Copyright Biomaterials.
50
51 1336x1871mm (95 x 95 DPI)
52
53
54
55
56
57
58
59
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28 Figure 3. Regulation of stem cell properties by controlling mechanical stress, temperature, and radiation in
29 bioprinting process. (A) Stress distribution in the needle during microextrusion-based bioprinting. Comparing
30 with static status, stress in flowing bioinks can lead to cell death near the shell of the needle. Reproduced
31 with permission from ref 109. Copyright Advanced Healthcare Materials. (B) A structure generated by
bioprinting of low-viscous methacrylated hyaluronic acid. In situ crosslinking makes it feasible to bioprint
32
low-viscous bioinks with high fidelity, which can decrease shear stress in microextrusion-based bioprinting.
33 Reproduced with permission from ref 117. Copyright Advanced Materials. (C) Viability of Chinese hamster
34 ovary cells after thermal inkjet bioprinting. Heat in thermal inkjet bioprinting caused cell death, which is
35 related to cell concentration. Reproduced with permission from ref 44. Copyright Biotechnology &
36 Bioengineering. (D) Spreading and viability of MSCs influenced by radiation in laser-assisted bioprinting.
37 Reproduced with permission from ref 77. Copyright Scientific Reports.
38
169x110mm (300 x 300 DPI)
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39 Figure 4. Microenvironmental factors modulate stem cell functions in long-term culture. (A) Bioink stiffness
40 affects osteogenic differentiation of MSC, stained by oil red O. Reproduced with permission from ref 129.
41 Copyright PloS one. (B) Fabricate surface nano-features to regulate cell morphology and migration.
Reproduced with permission from ref 130. Copyright Colloids and Surfaces B: Biointerfaces. (C) Bioprinting
42
hybrid bioactive structure of ECM to improve biocompatibility and PCL as the framework. Reproduced with
43 permission from ref 137. Copyright Nature Communications. (D) Co-printing of 10T1/2 fibroblasts (blue),
44 hNDFs (green), HUVECs (red) can facilitate the formation of heterogeneous tissue constructs by paracrine.
45 Reproduced with permission from ref 149. Copyright Advanced Materials. (E) Tethering RGD ligands onto ɑ-
46 cyclodextrin-modified PEG can enhance spreading of MSCs. Reproduced with permission from ref 152.
47 Copyright Advanced Materials. (F) Porous internal structure of alginate/gelatin hydrogel is essential for mass
48 transfer and cell migration. Reproduced with permission from ref 125. Copyright PloS one. (G) HUVECs (red)
printed in hNDF-laden (green) matrix form a linear vessel with the lumen to support perfusion for tissue
49 mature in long-term culture. Reproduced with permission from ref 165. Copyright Proceedings of the
50 National Academy of Sciences.
51
52 162x162mm (300 x 300 DPI)
53
54
55
56
57
58
59
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
Table 1. Comparison of bioprinting techniques
24
25 385x193mm (96 x 96 DPI)
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45 Table 2. Commonly used bioinks in stem cell bioprinting
46
47 529x776mm (72 x 72 DPI)
48
49
50
51
52
53
54
55
56
57
58
59

Bioprinting of Stem Cells Interplay of Bioprinting Process, Bioinks and Stem Cell Properties

Uploaded by

Document Information

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Bioprinting of Stem Cells Interplay of Bioprinting Process, Bioinks and Stem Cell Properties

Uploaded by

Copyright:

Available Formats

Subscriber access provided by TUFTS UNIV

is published by the American Chemical Society. 1155 Sixteenth Street N.W.,

13 bioprinting bioprinting bioprinting

14 Printing speed High Low Medium High Medium High

19 Cell density Low High Medium Medium High Low

20 Cost Low Low High High Medium High

25 References [20,22,34,35,44,102] [27,47-50,65,66,103] [16,70,71,104] [75,76,94,95] [86,90,94] [32,101,105,106]

7 Agarose/alginate/chitosan Micrextrusion-based Ionically NSCs >75 62

You might also like